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GENETIC RESOURCES,

CHROMOSOME ENGINEERING,

AND CROP IMPROVEMENT

Grain Legumes

EDITED BY

RAM J.SINGH AND PREM P.JAUHAR

Boca Raton London New York Singapore

A CRC title, part of the Taylor & Francis imprint, a member of

the Taylor & Francis Group, the academic division of T&F

Informa plc.

CHAPTER 7Chickpea (Cicer arietinum L.)F.Ahmad, P.M.Gaur, and J.S.Croser

7.1 INTRODUCTIONChickpea (Cicer arietinum L.), commonly called gram, Bengal gram, or garbanzo bean, is the most important food grain legume of South Asia and the third most important in the world after common bean (Phaseolus vulgaris L.) and field pea (Pisum sativum L.). Chickpea is a diploid with 2n=2x=l6 chromosomes and a genome size of approximately 750 Mbp (Arumuganathan and Earle, 1991). Chickpea is one of the first grain crops cultivated by man and has been uncovered in Middle Eastern archaeological sites dated to the eighth millennium BC (Zohary and Hopf, 2000). Two distinct market type classes, desi and kabuli, are recognized in chickpea (Pundir, Rao, and van der Maesen, 1985). The desi types that account for about 85% of chickpea area usually have small, angular-shaped, dark-colored seeds with a rough surface, pink flowers, anthocyanin pigmentation on the stems, and either semi-erect or semi-spreading growth habit. The kabuli type, which cover the remaining 15% area, usually have large “rams head”-shaped smooth surface seeds, lack of anthocyanin pigmentation, and semi-spreading growth habit. It has become increasingly clear during the last few decades that meeting the food needs of the world’s growing population depends, to a large extent, on the conservation and use of the world’s remaining plant genetic resources. Conservation without use has little point and use will not come without evaluation. Genetic resources encompass all forms of the cultivated species, as well as their related wild species (Harlan, 1984). That is a general concept to which chickpea is no exception. In reviewing genetic resources and their multifaceted applications in chickpea genetic improvement, we have placed more emphasis on the wild genetic resources of the cultivated chickpea, while providing a brief overview of resources available in the cultivated species.

7.1.1 Global Chickpea Production and DistributionChickpea is grown in more than 40 countries, but the main growing region is South and Southeastern Asia (ca. 70%), where India (ca. 60%) and Pakistan (ca. 10 to 15%) are major contributors. West Asia accounts for approximately 16% of global chickpea area, with Iran, Turkey, and Syria being the largest producers in the region. Africa accounts for

5%, mostly from Ethiopia, Malawi, and Tanzania in Eastern Africa and Morocco in North Africa. In remaining parts of the world, about 3% area is contributed by North America (mainly Canada), 2% by Australia, and 1% by Europe (mainly Spain). The global chickpea area has remained almost stagnant during the past 40 years; it was 11.8 m/ha during 1961–1965 and 11.0 m/ha during 1996–2000. Though there has not been much change in the global chickpea area, the production has increased from 7 m/t in 1961–1965 to 8.4 m/t during 1996–2000, due to enhancement in yield levels from 599 to 791 kg/ha 1 during this period. During 2000–2002, chickpea production was 7.5 m/t from an area of 9.7 m/ha (average of three years). There has been more than a sevenfold increase in the global trade of chickpea during the past four decades. Trade increased from 0.13 m/t during 1961–1965 to 0.66 m/t during 1996–2000 and reached its highest level of 1 m/t during 2001. India has been the world’s largest chickpea importing country during the past 15 years. It had a record import of 0.52 m/t during 2001. Pakistan has emerged as the second largest importer of chickpea during this period, its imports increasing from 5,000 t during 1981–1985 to 0.1 m/t during 2001. Other countries that have had sizable chickpea import during recent years are Spain, Saudi Arabia, Italy, Jordan, Tunisia, Lebanon, Turkey, Sri Lanka, and Colombia (Gowda and Gaur, 2004). 7.1.2 Importance of the CropChickpea has one of the highest nutritional compositions of any dry edible grain legume and does not contain significant quantities of any specific major antinutritional factors. On an average, chickpea seed contains 23% of highly digestible protein, 64% total carbohydrates, 47% starch, 5% fat (primarily linoleic and oleic acids), 6% crude fiber, 6% soluble sugar, and 3% ash. The mineral component is high in phosphorus (343 mg/100 g), calcium (186 mg/100 g), magnesium (141 mg/100 g), iron (7 mg/100 g), and zinc (3 mg/100 g) (Williams and Singh, 1987). Used in a variety of ways, chickpea is not only good for human health but also for soil health. It meets 80% of its nitrogen (N) requirement from a symbiotic rhizobial interaction, which enables the crop to fix up to 140 kg N ha 1 from atmosphere (Saraf et al., 1998). It leaves substantial amount of residual nitrogen behind for subsequent crops and adds much needed organic matter to maintain and improve soil health, long-term fertility, and sustainability of the ecosystems. In recent years, chickpea has also gained popularity in broad-acre cropping systems in developed countries, particularly Australia and Canada (Siddique and Sykes, 1997). A dryland crop requiring minimal inputs, chickpea is a boon to the resource-poor marginal farmers in the tropics.

Genetic resources, chromosome engineering, and crop improvement 230

7.2 ORIGIN OF CROP AND ITS TAXONOMY7.2.1 Origin of ChickpeaThe available evidence suggests that chickpea originated in the fertile crescent region of southeastern Turkey and adjoining Syria (van der Maesen, 1987). The proposed wild progenitor and several other annual Cicer species occur there (Ladizinsky, 1975). Further evidence of its origin comes from seeds dated to about 5450 BC, unearthed from archaeological excavations at Hacilar near Burdur in Turkey (Helbaek, 1970). It is believed that chickpea diverged from Turkey in two directions—into the western parts where it is grown in the spring and summer and into the eastern and southern parts where it is grown in the cool dry seasons. The majority of the wild Cicer species are found in the West Asia and North Africa region covering Turkey in the north to Ethiopia in the south, and Pakistan in the east to Morocco in the west. Botanically, the cultivated chickpea has been split into two groupings, microsperma and macrosperma,corresponding to seed size and in much the same way as it has been done for lentils (Cubero, 1987). From a practical point of view, chickpea is also classified into kabuli and

desi types. The terms desi and kabuli, however, do not overlap with microsperma and macrosperma. The kabuli types are now grown predominantly in the countries of the Mediterranean region, West Asia, North Africa, Australia, and North America, while the desi types are grown mostly in South Asia, Iran, Ethiopia, Mexico, and Australia. Other than morphological differences, all forms of the cultivated species share the same genome. The wild annual progenitor of chickpea has been repeatedly identified as the annual species C. reticulatum Lad. (Ladizinsky and Adler, 1976b; Ahmad, Gaur, and Slinkard, 1992; Iruela et al., 2002). It is believed that one of the crucial steps in the origin of cultivated chickpea was the change from the perennial to annual life cycle (Gupta and Bahl, 1983). The perennial species have not been extensively studied, however, the available evidence suggests C. anatolicum Alef. to be the probable perennial progenitor of C. arietinum (Gupta and Bahl, 1983; Ahmad, 1989; Tayyar and Waines, 1996). It appears that domestication and, thus, evolution of the cultivated species followed the usual process of artificial selection, which favored large palatable seeds, reduced pod dehiscence, nondormancy, synchronous ripening, earliness, and diversity of forms (van der Maesen, 1987).

7.2.2 Taxonomy of CicerThe cultivated chickpea species has been taxonomically placed in the genus Cicer, which belongs to the family Fabaceae and its monogeneric tribe Cicereae Alef. (Kupicha, 1981). Presently, the genus Cicer consists of 43 species (Table 7.1), divided into 4 sections, Monocicer, Chamaecicer, Polycicer, and Acanthocicer, based on their morphological characteristics, life cycle, and geographical distribution (van der Maesen, 1987; http://singer.cgiar.org/Search/SINGER/search.htm). Eight

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Table 7.1 List of All Known Species in the GenusCicer

Annual SpeciesC. arietinum* L. C. judaicum* Boiss. C. bijugum* K.H.Rech. C. pinnatifidum* Jaub. & Sp. C. chorassanicum* (Bge.) M.Pop. C. reticulatum* Ladiz. C. cuneatum* Hochst. ex Rich. C. yamashitae* Kitamura C. echinospermum* P.H.Davis Perennial SpeciesC. acanthophyllum Boriss. C. macracanthum M.Pop. C. anatolicum* Alef. C. microphyllum* Benth. C. atlanticum Coss. ex Maire C. mogoltavicum (M. Pop.) Koroleva C. balcaricum Galushko C. montbretii* Jaub. & Sp. C. baldshuanicum (M.Pop.) Lincz. C. multijugum van der Maesen C. canariense* Santos Guerra & Lewis C. nuristanicum Kitamura C. fedtschenkoi Lincz. C. oxyodon Boiss. & Hoh. C. flexuosum Lipsky C. paucijugum (M. Pop.) Nevski C. floribundum* Fenzl. C. pungens* Boiss. C. graecum Orph. C. rassuloviae Lincz. C. grande (M. Pop.) Korotk. C. rechingeri Podlech C. heterophyllum* Contandr et al. C. songaricum* Steph. ex. DC. C. incanum Korotk. C. spiroceras Jaub. & Sp. C. incisum* (Willd.) K.Maly C. stapfianum K.H.Rech. C. isauricum* P.H.Davis C. subaphyllum Boiss. C. kermanense Bornm. C. tragacanthoides Jaub. & Sp. C. korshinskyi Lincz. UnspecifiedC. laetum Rassulova & Sharipova *Species with confirmed somatic chromosome number of 2n=16 Source: The CGIAR System-Wide Information Network for Genetic Resources (SINGER; http://singer.cgiar.org/Search/SINGER/search.htm) and van der Maesen, 1987. Adapted from Croser et al. 2003a.


of these Cicer species, sharing the annual growth habit with chickpea, are of particular interest to breeders. Of the nine annual Cicer species, eight are classified within the Monocicer section, and one, C. chorassanicum, within the Chamaecicer section (Kazan and Muehlbauer, 1991; Muehlbauer, Kaiser, and Simon, 1994). Thirty-three of the remainder species are known to be perennial, while C. laetum Rass. & Sharip has an unspecified life cycle (van der Maesen, 1987).

7.3 GERMPLASM COLLECTION, MAINTENANCE, AND EVALUATIONDue to its importance as an affordable protein source in developing nations, it is crucial to obtain and maintain higher yields of the chickpea crop. Effective genetic means are, thus, needed to reach such key objectives. The effectiveness of selection in any crop depends upon the extent and nature of phenotypic and genotypic variability present in different traits of the population (Arora, 1991). Unfortunately, the chickpea crop is susceptible to a range of biotic and abiotic stresses, which can be devastating to crop yield. A low level of genetic variability within C. arietinum has hampered chickpea breeders in their efforts to develop widely adapted cultivars with resistance to biotic and abiotic stresses (Ladizinsky and Adler, 1975; Tuwafe et al., 1988; Kazan and Muehlbauer, 1991; Ahmad et al., 1992; Ahmad and Slinkard, 1992; Udupa et al., 1993; van Rheenen et al., 1993; Labdi et al.,1996; Tayyar and Waines, 1996; Simon and Muehlbauer, 1997; Siddique et al., 2000). Recently, this low level of genetic variability has been attributed to a series of genetic bottlenecks in the domestication of chickpea, including the restricted distribution of the wild progenitor, the founder effect associated with domestication, and the shift from winter to summer cropping (Abbo et al., 2003). To compound this problem, genetic erosion of C. arietinum intraspecific resources is occurring due to the loss of local ecotypes, through diseases, insects, and environmental stresses, as well as for economic and strategic reasons (Malhotra et al., 2000; Abbo et al., 2003). Breeders are therefore looking to the wild relatives of chickpea as an alternative genetic resource for crop improvement. The world collection of annual wild Cicer species is very limited and currently consists of 593 entries held in nine genebanks around the world. These represent only 285 known separate accessions of the eight annual wild Cicer species (Berger et al., 2003). Of these, only 120 have been collected independently from wild populations, with the remainder being selections from the original material. There is a single original accession of C. cuneatum, 2 of C. chorassanicum, 13 of C. echinospermum, 18 of C.

reticulatum, 21 of C. bijugum, 28 of C. pinnatifidum, three of C. yamashitae, and 34 of C.

judaicum. Thus, only a small proportion of the habitat and genetic diversity that is potentially available in wild populations is present in ex situ collections. It is essential to widen genetic diversity of the world collection of annual wild Cicer species by conducting targeted missions, which address the multitude of gaps in the current collection (Berger et al., 2003). The world germplasm collection of perennial Cicer species is even more limited. Compared to the annual species, screening has been difficult due to the inherent problems with growing these species so far from their adapted climatic conditions. Malhotra et al.

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(2000) have suggested that they should be conserved in situ by preserving their original habitats. Recent progress in determining adequate growing conditions for some of the perennial species, and subsequent establishment of perennial Cicer nurseries in eastern Washington state should go some way to improving this situation in the future (Kaiser et al., 1997). Approximately 12 of the 34 perennial species have been maintained, with various degrees of success, at The Germplasm Resources Information Network-United States Department of Agriculture (Pullman, WA). 7.4 TRAITS OF ECONOMIC SIGNIFICANCE WITHIN THE WILD CICER GENE POOL

Comprehensive screening of wild Cicer collections at the International Centre for Agricultural Research in Dry Areas (ICARDA), Syria, the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), India, and other national institutions have identified germplasm with resistance to various diseases and insects. Table 7.2 lists the traits of economic significance currently identified within the wild annual and perennial species of Cicer.Wild Cicer species are the only source of resistance so far found for bruchid (Callosobruchus chinesis L.) and cyst nematode. They have a higher level of resistance than the cultivated species for fusarium wilt, leaf miner, phytophthora root rot, drought, ascochyta blight, pea streak carlavirus, and cold. Perhaps even more importantly, several accessions of the wild Cicer species are resistant to three or more stresses (Robertson et al., 1996). Particularly promising are the following accessions that are resistant to four or five different stresses, viz. C. reticulatum (ILWC 81, 112), C. echinospermum (ILWC 39, 181), C. bijugum (ILWC 32, 62, 73, 79), C. judaicum (ILWC 46), and C. pinnatifidum(ILWC 236). Such accessions are obvious targets for interspecific hybridization efforts. Overall, the species containing accessions with the highest levels of resistance to the most stresses in order of performance are: C. bijugum, C. pinnatifidum, and C. judaicum(Singh et al., 1994; Singh et al., 1998). Accessions of the eight wild annual Cicer species have also been evaluated for various morphological traits at ICARDA and a catalogue was prepared (Robertson et al., 1995). Robertson et al. (1997) have reported useful variations for vegetative and reproductive characters in the annual wild species. Not surprisingly, C. arietinum showed greater intraspecific morphological variability compared with the wild species, particularly for characters like leaf area, growth habit, plant height, first pod height, pod dehiscence, and 100-seed weight. Among the wild species, C. reticulatum showed the largest overall morphological variability, followed by C. pinnatifidum, C. echinosper-


Table 7.2 Desirable Traits Identified in Wild Annual and Perennial Cicer SpeciesTrait Species with Resistance/Tolerance Reference(s)Ascochyta blight C. anatolicum, C. bijugum, C. cuneatum, C.

echinospermum, C. judaicum, C. montbretti,

C. pinnatifidum, C. reticulatum Haware et al., 1992; Singh and Reddy, 1993; Stamigna et al., 1998; Singh et al., 1998; Collard et al., 2001; Collard et al., 2003a Botrytis grey mould C. bijugum Haware et al., 1992

Bruchid C. bijugum, C. cuneatum, C. echinospermum, Singh et al., 1994; Singh et al., 1998C. judaicum, C. pinnatifidum, C. reticulatum Cold C. bijugum, C. echinospermum, C. judaicum,

C. microphyllum, C. pinnatifidum, C.

reticulatum

van der Maesen and Pundir, 1984; Singh et al., 1990,1995; Singh et al., 1998; Toker, 2004 Cyst nematode C. bijugum, C. pinnatifidum, C. reticulatum Singh et al., 1989; Singh and Reddy, 1991; Singh et al., 1994, 1996; Singh et al., 1998; Di Vito et al., 1996 Crenatebroomrape C. bijugum, C. canariensis, C.

echinospermum, C. judaicum, C.

macracanthum, C. multijugum, C. oxyodon, C.

pinnatifidum, C. reticulatum, C. songaricum,

C. yamashitae

Rubiales et al., 2004

C. bijugum, C. echinospermum, C.

judaicum, C. pinnatifidum, C.

reticulatum C. bijugum, C.

canariense, C. cuneatum,Drought C. microphyllum Chandel, 1984 .... implied based on distribution Fusarium wilt C. bijugum, C. echinospermum, C. judaicum,

C. pinnatifidum, C. reticulatum

Nene and Haware, 1980; Haware et al., 1992; Singh et al., 1994; Kaiser et al., 1994; Infanito et al., 1996; Porta-Puglia and Infantino, 1997Helocoverpapod borer C. bijugum, C. canariense, C. cuneatum, C.

echinospermum, C. judaicum, C.

maracanthum C microphyllum C

Kaur et al., 1999, Sharma et al., 2002, 2004

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pinnatifidum, C. reticulatum Leaf miner C. bijugum, C. chorassanicum, C. cuneatum,

C. echinospermum, C. judaicum, C.

pinnatifidum, C. reticulatum

Singh and Weigand, 1994; Singh et al., 1994, 1998 Pea streak carlavirus C. anatolicum, C. canariensis, C.

microphyllum, C. oxyodon

Kaiser et al., 1993 Phytophthora root rot C. echinospermum Singh et al., 1994; Knights et al., 2003Trypsin inhibitor C. chorassanicum Ahmad and Kollipara, 2004 mum, and C. bijugum. Of interest was the variability for wide leaflets in C.

chorassanicum, number of branches in C. bijugum, and C. reticulatum and early flowering in C. judaicum.In terms of nutritional value, there is a wide variation in seed protein and amino acid content in the annual Cicer species (Singh and Pundir, 1991). Seed protein ranges from 168 g/kg in C. cuneatum to 268 g/kg in C. pinnatifidum. Despite this, prospects for upgrading the nutritional value of chickpea by introgression of genes for high protein content are low, as the variability in the wild accessions falls within the range for the cultivated chickpea (Ocampo et al., 1998). Further, chickpea seed extract is known to inhibit trypsin and chymotrypsin (Saini et al., 1992), however, preliminary data for trypsin protease inhibitor indicate that there is no such activity in C. chorassanicum(Ahmad and Kollipara, 2004). 7.5 SPECIES RELATIONSHIPS AND INTERSPECIFIC HYBRIDIZATION

7.5.1 Species Relationships within the GenusTraditionally phenotypic traits (Nozzolillo, 1985; De Leonardis et al., 1996; Robertson et al., 1997; Hassan, 2000; Javedi and Yamaguchi, 2004a), hybridization success (Ladizinsky and Adler, 1976a, 1976b; Pundir and van der Maesen, 1983; Verma et al., 1990; Pundir et al., 1992; Sheila et al., 1992; Pundir and Mengesha, 1995; Badami et al., 1997; Singh et al., 1999a, 1999b; Stamigna et al., 2000), analysis of chromosome pairing in hybrids (Ladizinsky and Adler, 1976a, 1976b; Ahmad et al., 1987; Ahmad, 1988), and the study of chromosome structure (Ohri and Pal, 1991; Ocampo et al., 1992; Tayyar et al., 1994; Ahmad, 2000) have been widely used methods for analysis of genomic relationships and the construction of phylogenies among Cicer species. Over the past 15 years, electrophoretic data based on seed storage protein (Ladizinsky and Adler, 1975; Vairinhos and Murray, 1983; Ahmad and Slinkard, 1992) and isozymes (Kazan and


Muehlbauer, 1991; Ahmad et al., 1992; Labdi et al., 1996; Tayyar and Waines, 1996; Gargav and Gaur, 2001, Sudupak and Kence, 2004) have also been applied to systematic studies in Cicer. More recently, DNA-based techniques (Patil et al., 1995; Sharma et al., 1995; Ahmad, 1999; Choumane et al., 2000; Iruela et al., 2002; Sudupak et al., 2002; Rajesh et al., 2003; Javedi and Yamaguchi, 2004a, b; Nguyen et al., 2004; Sudupak, 2004; Sudupak et al., 2004) have provided many new approaches to compare aspects of genome relationships in ways not previously possible. Among the annual Cicer species, there is a consensus that C. reticulatum and C.

echinospermum are the wild species most closely related to the domesticated C.

arietinum. Cicer bijugum, C. pinnatifidum, and C. judaicum show a closer relationship among themselves and appear to form a group that is next closest to the first group containing the cultivated species. The remainder of the annual species, viz. C.

chorassanicum, C. yamashitae, and C. cuneatum share an even more distant relationship with the cultivated species. This relationship only generally holds true, as the five annual species that are not part of the group with the cultivated species appear to change positions for relatedness to each other and to C. arietinum. Perennial Cicer species have only recently been subjected to phylogenetic analysis (Tayyar and Waines, 1996; Gargav and Gaur, 2001; Iruela et al., 2002; Sudupak et al., 2002; Rajesh et al., 2003; Javedi and Yamaguchi, 2004a, b; Nguyen et al., 2004; Sudupak, 2004; Sudupak and Kence, 2004; Sudupak et al, 2004). Again, with some exceptions, the perennial species in general have shown a distant relationship and are far removed from the cultivated species. The relationship between the perennial species C. anatolicum and domesticated chickpea remains a contentious issue, with some studies placing this species as a close relative of the cultivated species, while others indicating it to be far removed (Kazan and Muehlbauer, 1991; Staginnus et al., 1999; Choumane et al., 2000; Rajesh et al., 2003; Nguyen et al., 2004; Sudupak, 2004; Sudupak and Kence, 2004). 7.5.2 Gene Pools and Interspecific HybridizationLadizinsky and Adler (1976a, 1976b) and van der Maesen (1987) adopted the classical definition of the primary, secondary, and tertiary gene pools as proposed by Harlan and de Wet (1971) for classification of the Cicer wild relatives. According to this definition, the primary gene pool includes all species that hybridize freely, show good chromosome pairing leading to gene exchange, and produce viable hybrids. The secondary gene pool includes species that can be used as germplasm resources; however, hybridization with the cultivated species is difficult because of genetic barriers or chromosome alterations, and some degree of sterility is associated with the first-generation hybrids. Members of the tertiary gene pool are difficult to utilize, and sterility is always associated with hybrids. Fertility can sometimes be restored, but usually the percentage of recovered viable zygotes is extremely small. On the basis of Harlan and de Wet’s (1971) definition, and in consideration of the results obtained from crossability, biochemical and molecular diversity, and karyotypic studies, a recently revised model of the wild annual Cicer gene pools has been proposed (Croser et al., 2003a). However, if one were to follow Harlan and de Wet’s (1971) definition alone, the primary gene pool of Cicer would consist of C. arietinum and only one wild species, the wild annual progenitor C. reticulatum. Although the other closely

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related species, C. echinospermum, crosses quite readily with the cultivated species, it shows varying levels of sterility in the F1 and F2 generations. The secondary gene pool, thus, consists of C. echinospermum only. Cicer bijugum, C. pinnatifidum, and C.

judaicum, which have been reported to give hybrids when crossed with the cultivated species (Verma et al., 1990; Singh et al., 1994; Singh et al., 1999a, 1999b), have been placed in the secondary gene pool by Croser et al. (2003b). This, however, is an issue that is not resolved yet and calls for some discussion. Although successful crosses of C.

bijugum, C. pinnatifidum, and C. judaicum with C. arietinum have been reported, the authors have as yet failed to produce (except the C. arietinum×C. judaicum hybrid) (Verma et al., 1995) conclusive proof of hybridity by any cytogenetical, biochemical, or DNA-based approach. Indeed, there is a growing skepticism in the scientific community about the authenticity of these apparent partially fertile hybrids and their resulting progenies (Pundir, personal communication, 1994; Cubero and Ocampo, personal communication, 2004; Muehlbauer, personal communication, 2004). Considering the ongoing scientific dilemma and the lack of success, despite repeated attempts by various researchers globally utilizing embryo and ovule culture procedures and spanning more than two decades, we conclude that partially fertile interspecific hybrids and their progenies as claimed by the above research groups (Verma et al., 1990; Singh et al, 1994; Singh et al., 1999a, 1999b) are highly questionable. We would, thus, propose that the above three species should be placed in the tertiary gene pool of chickpea, along with the remaining annual species C. chorassanicum, C. yamashitae, and C. cuneatum. The lack of availability of perennial Cicer species has greatly restricted the assessment of their crossability with the cultivated species. Given the current situation with perennial Cicerspecies (Croser et al., 2003a), and until proven otherwise, we feel these species should be appropriately placed in the tertiary gene pool along with the six other annual wild species.It is obvious from crossability studies of the cultivated species with the wild Cicerspecies that technical difficulties in obtaining hybrids beyond those within the primary and secondary gene pools remain a major obstacle. Apart from a few rather surprising exceptions (Verma et al., 1990; Singh et al., 1994; Singh et al., 1999a, 1999b), postfertilization incompatibility barriers (Bassiri et al., 1987; Ahmad et al., 1988; Stamigna et al., 2000; Ahmad and Slinkard, 2004) have restricted successful hybridization (using conventional crossing techniques) exclusively to the species of the primary and secondary gene pools. Given the fact that the tertiary gene pool species remain crossincompatible with the cultivated species, bridging crosses deserve further attention in chickpea in light of the known close relationships of C. arietinum with the wild annual species C. reticulatum and C. echinospermum. Ahmad and Slinkard (2003) have recently shown that, provided reciprocal crosses are used, there are at least no prefertilization barrier(s) limiting the hybridization of C. reticulatum and C.

echinospermum with the other annual wild Cicer species. If the above-mentioned three species can be crossed with any one of the wild annual or perennial species, then it would be possible to transfer traits of interest to C. arietinum.Rescuing abortive interspecific embryos and their in vitro culture has been proposed as an effective strategy for overcoming species barriers (van Rheenen, 1991; Muehlbauer et al., 1994; Singh et al. 1994; Robertson et al., 1996; Singh and Ocampo, 1997). In Cicer, there has been limited success from attempts to develop tissue culture techniques


to enable hybridization between chickpea and the more distantly related annual wild species C. cuneatum, C. pinnatifidum, and C. bijugum (Singh and Singh, 1989; Swamy and Khanna, 1991; Verma et al., 1995; Badami et al., 1997; van Dorrestein et al., 1998; Mallikarjuna, 1999, 2001). To date, a technique that enables the reproducible rescue and growth of embryos before eight days after pollination is lacking in Cicer. Thus, there is a clear need to identify and fine-tune information regarding the physiological, nutritional, and hormonal requirements of developing chickpea embryos (Croser, 2002; Croser et al., 2003a). An international collaboration has been established between Canadian, Australian, and Indian researchers to develop techniques for efficient interspecific hybridization between the cultivated and wild Cicer species. 7.6 CYTOGENETICS AND LINKAGE MAPS OF CHICKPEA

7.6.1 CytogeneticsChickpea is a crop that is not amenable to cytogenetic studies. Hence, most studies are limited to chromosome counts and feulgen-stained studies of karyotypes because of the small size and sticky nature of chromosomes. No cytogenetic stocks, other than tetraploids, are available in chickpea (Bahl, 1987; Gupta and Sharma, 1991) and except for one (Vlacilova et al., 2002), linkage groups have not yet been associated with respective chromosomes. Cicer arietinum has been the subject of a considerable number of karyotypic studies, while the wild annual species have received less attention (Croser et al., 2003a). The nine annual and nine of the remainder 34 species are confirmed diploids with 2n=2x=16 chromosomes (Table 7.1) (Ladizinsky and Adler, 1976a, 1976b; Ahmad, 1989; Pundir et al., 1993; Ohri, 1999; Ahmad, 2000; Ahmad and Chen, 2000). The karyotypes of the primary gene pool species C. arietinum, C. reticulatum, C.

echinospermum, and the tertiary gene pool species C. songaricum and C. anatolicumshow a high degree of similarity to each other and form the first group, while the second group contains all of the remaining six annual species (Ahmad, 2000; Croser et al., 2003a). Karyotypic details of the other species in the genus are not known. The crossability of these two perennial species with C. arietinum remains largely unknown. Whether a karyotypic similarity-crossability equation holds true in the Cicer genus needs additional research, but broad karyotypic and crossability studies of the cultivated chickpea with perennial Cicer species for alien gene introgression appear warranted. Our understanding of the cytogenetic map in chickpea and related Cicer species is very much lacking. Individual chromosomes of chickpea are reported to be identifiable by C-banding and fluorochrome staining (Galasso and Pignone, 1992; Tayyar et al., 1994; Galasso et al., 1996). The related annual wild species have also been subjected to a solitary C-banding analysis (Tayyar et al., 1994), and the general conclusion drawn is that the heterochromatic C-bands are located proximally around the centromere with only occasional bands in intercalary and distal positions. Pachytene chromosome analysis of the cultivated species (Ahmad and Hymowitz, 1993) has further corroborated such distribution of heterochromatin. While the authors (Tayyar et al., 1994) were able to

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identify individual pairs of chromosomes within a species, its applicability in an interspecific hybrid situation remains to be explored. Karyotype analyses have shown that only one chromosome pair is associated with the nucleolar organizing regions, and is thus considered satellited (Ocampo et al., 1992; Ahmad, 2000). The satellite is present on the longest first chromosome in C. arietinum,

C. reticulatum, and C. echinospermum, and an extra satellite in the case of C. reticulatummay also be present on the second chromosome. Unlike the somatic karyotype, detailed pachytene chromosome analysis had shown the third chromosome to be clearly satellited in C. arietinum (Ahmad and Hymowitz, 1993). Such refined pachytene analysis has made individual chromosome identification more conclusive and adds a further dimension to chickpea cytology (Ahmad and Hymowitz, 1993). Given the poor state of chickpea cytology, there has been commendable progress in molecular cytogenetics in the genus. Individual chickpea chromosomes have been successfully sorted by flow cytometry (Vlacilova et al., 2002) and utilized for mapping specific DNA sequences and genes to individual chromosomes. Thus, specific genes (coding for various rRNA loci), major random repetitive DNA sequences, STMS markers, microsatellites, En/Spm-like transposon sequences, simple sequence repeats, and Arabidopsis-type telomeric sequences have been successfully hybridized to and localized on the chickpea chromosomes by fluorescent in situ hybridization (FISH) (Abbo et al., 1994; Galasso et al., 1996; Gortner et al., 1998; Staginnus et al., 1999, 2001; Vlacilova et al., 2002; Valarik et al., 2004). Using polymerase chain reaction and FISH, Vlacilova et al. (2002) have successfully associated two STMS markers (belonging to linkage group 8 of Winter et al., 2000) to the shortest chromosome of the chickpea genome. More recently, FISH analysis on super-stretched flow-sorted chickpea chromosomes has revealed spatial resolution of neighboring loci that has not been obtained by any other method (Valarik et al., 2004). It is expected that further technical advances will lead to the development in genome mapping of chickpea and the association of all genetic linkage groups to specific well-defined chromosomes. 7.6.2 Genetic Linkage MappingDevelopment of high-density integrated genetic linkage maps based on morphological, biochemical, and DNA markers is a prerequisite for use in marker-assisted selection, positional cloning, and mapping of quantitative trait loci of agronomically important traits in a crop species. Although an impressive amount of progress has been achieved in linkage mapping, chickpea researchers are still awaiting a sufficiently dense genetic linkage map. Intraspecies polymorphism in cultivated chickpea for commonly used molecular markers is extremely low, therefore, interspecific crosses (C. arietinum×C.

reticulatum, C. arietinum×C. echinospermum) have been exploited to develop genetic linkage maps with a higher number of markers (Gaur and Slinkard, 1990a, 1990b; Simon and Muehlbaur, 1997; Winter et al., 2000; Tekeoglu et al., 2002; Pfaff and Kahl, 2003). A genetic map constructed from an interspecific cross may not represent the true recombination distance map order of the cultivated genome due to uneven recombination of homoeologous chromosomes and distorted genetic segregation ratios (Flandez-Galvez et al., 2003a). A genetic linkage map constructed from a cross within the cultivated gene pool (Cho et al., 2002; Flandez-Galvez et al., 2003a), especially in the framework of


targeting traits of breeding importance, would therefore be more desirable. The genetic linkage maps developed to date with morphological, biochemical, and DNA-based molecular markers are summarized in Table 7.3. The beginnings of linkage map development in chickpea were based on morphological and isozyme loci. However, their small numbers and the fact that expression of these markers is often influenced by the environment, makes them unsuitable for routine use. Thus, these maps were sparse and represented less than 30 loci mapped in a very small portion (about 250 cM) of the chickpea genome (Gaur and Slinkard, 1990a, 1990b; Kazan et al., 1993). Later developments on genetic linkage maps in chickpea that started with the work of Simon and Muehlbauer (1997) relied heavily on DNA-based molecular markers. In the lack of more recently available molecular markers, Simon and Muehlbauer (1997) employed RFLP and RAPD markers that show limited polymorphism in the cultivated species (Udupa et al., 1993; Banerjee et al., 1999). Two independent interspecific-derived populations have been extensively employed for genetic linkage map development in chickpea (C. arietinum ICC 4958×C. reticulatumPI 489777 at the University of Frankfurt, Germany, and C. arietinum FLIP 84–92C×C.

reticulatum PI 599072 at Washington State University, Pullman, WA), with the one developed in Germany being denser than the other. The 982 cM distance linkage map developed by Santra et al. (2000), which consisted of 89 RAPD, 17 ISSR, 9 isozyme, and 1 morphological markers was further expanded (Rajesh et al., 2002b; Tekeoglu et al., 2002) by integrating sequence tagged microsatellite sites (STMS) and resistance gene analog (RGA) loci. It covers a distance of 1175 cM. The STMS markers have proved very useful in linkage mapping and formed the basis for the map initially developed by Winter et al. (1999) that spanned a distance of 613 cM and consisted of 120 STMS markers. This map was greatly extended by Winter et al. (2000) and more recently by Pfaff and Kahl (2003) with the addition of 47 defense response (DR) genes. It covers a distance of 2500 cM arranged in 12 linkage groups and represents the most extensive linkage map in chickpea (Table 7.3). Relatively smaller maps derived from intraspecific (within C. arietinum) crosses, are also being developed (Cho et al., 2002; Flandez-Galvez et al., 2003a). Combining these maps into a consensus linkage map is in progress and is being facilitated by the International Chickpea Genomics Consortium (Rajesh et al., 2004; http://www.icgc.wsu.edu/). Although steady progress is being made in extending the genetic linkage map in chickpea, there has been little effort in relating these maps to the eight pairs of chromosomes in the cultivated chickpea. Using flow-sorted chromosomes and molecular cytogenetic analysis, a recent report (Vlacilova et al., 2002) has linked STMS markers GAA46 and TS45 (linkage group 8 of the linkage map developed by Winter et al., 2000) to chromosome number 8 of chickpea.

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Table 7.3 Genetic Resources Utilized in the Evolution of Genetic Linkage Map Development in Chickpea*Publication Mapping Population Remarks (map size, markers)Gaur and Slinkard, 1990a, F2, intraspecific (C. reticulatum) 200 cM, 7 linkage groups

1990b F2, interspecific (C. arietinum×C.

reticulatum) and F2, interspecific (C. arietinum×C. echinospermum)

3 morphological and 26 isozymes Kazan et al., 1993 F2, intraspecific (C. arietinum) 257 cM, 8 linkage groups F2, interspecific (C. arietinum×C.


5 morphological and 23 isozymes Simon and Muehlbauer, 1997

F2, interspecific (C. arietinum×C.


550 cM, 10 linkage groups 9 morphological, 27 isozyme, 10 RFLP and 45 RAPD Winter et al., 1999 RIL, interspecific (C. arietinum×C.

reticulatum)

613 cM, 11 linkage groups 120 STMS Winter et al., 2000 Same as Winter et al.,1999 2078 cM, 16 linkage groups 118 STMS, 96 DAF, 70 AFLP, 37 ISSR, 17 RAPD, 8 isozyme, 3 cDNA, 2 SCAR and 3 morphological Santra et al., 2000 RIL, interspecific (C. arietinum×C.

reticulatum)

982 cM, 9 linkage groups 89 RAPD, 17 ISSR, 9 isozyme, and 1 morphological Hajj-Moussa et al., 2001 RIL, interspecific (C. arietinum×C.

reticulatum)

23 linkage groups RAPD, ISSR, and morphologicalRajesh et al., 2002b Same as Santra et al., 2000 Addition of RGA Potkin 1–2 171 to Linkage group 5 of Santra et al. (2000) Tekeoglu et al., 2002 Same as Santra et al., 2000 Extended map of Santra et al. (2000) to 1,175 cM


50 STMS and 1 RGA was integrated Cho et al., 2002 RIL, intraspecific (C. arietinum) 297 cM, 14 linkage groups 68 STMS, 34 RAPD, 4 ISSR, and 5 morphological Pfaff and Kahl, 2003 Same as Winter et al. 1999; 2000 Incorporated 47 DR gene specific markers to Winter et al. (2000) 2500 cM, 12 linkage groups Flandez-Galvez et al., 2003a F2, intraspecific (C. arietinum) 535 cM, 8 linkage groups 51 STMS, 3 ISSR, 12 RGA Collard et al., 2003b F2, interspecific (C. arietinum×C.

echinospermum)

570 cM, 8 linkage groups 14 STMS, 54 RAPD, 9 ISSR, 6 RGA * RIL: Recombinant Inbred Line, RFLP: Restriction Fragment Length Polymorphism, RAPD: Random Amplified Polymorphic DNA, STMS: Sequence Tagged Microsatellite Site, DAF: DNA Amplification Fingerprint, AFLP: Amplified Fragment Length Polymorphism, SCAR: Sequence Characterized Amplified Region, ISSR: Inter Simple Sequence Repeat, RGA: Resistance Gene Analog, DR: Defense Response. 7.7 GERMPLASM ENHANCEMENT

7.7.1 International Efforts in Chickpea ResearchSystematic international efforts on chickpea research started after the establishment of two future harvest Centers of Consultative Group on International Agricultural Research (CGIAR). The International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) was established in 1972, with its headquarters in Patancheru, India, and the International Center for Agricultural Research in the Dry Areas (ICARDA) was established in 1977, with its headquarters in Aleppo, Syria. ICRISAT works on both desiand kabuli chickpeas, while ICARDA concentrates on kabuli chickpea. In collaboration with National Agricultural Research Systems (NARS), these centers have made extensive efforts on collection, characterization, and conservation of chickpea germplasm. The genebank of ICRISAT has more than 17,000 accessions of the cultivated species and 136 accessions of 18 wild species; similarly the genebank of ICARDA has more than 12,000 accessions of the cultivated species and 260 accessions of 8 wild species. These precious assets held, in trust for humanity, by these centers are provided freely to research and development specialists around the world. More than 116,000 seed samples of germplasm accessions from ICRISAT and more than 19,000 from ICARDA have been disseminated to requesting NARS. Given the large number of accessions available in the genetic resource collection, it becomes a daunting task to identify germplasm that could be used in a crop improvement program. A chickpea core collection, which is a chosen subset of large germplasm collection that generally contains about 10% of the total accessions and represents the genetic variability of the entire germplasm collection, as

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recently developed by Upadhyaya and Ortiz (2001) and Upadhyaya et al. (2001), should greatly facilitate global chickpea breeding and genetic improvement. Fortunately, in chickpea, 15 genebank accessions supplied by ICRISAT and 17 supplied by ICARDA have been directly released as varieties by national authorities in different countries. Additionally, ICRISAT and ICARDA have been helping the chickpea breeding programs of NARS around the world by making available segregating materials and advanced breeding lines. A total of 50 varieties released in 10 countries are from ICRISAT-supplied breeding materials. Similarly, 97 varieties in 25 countries are from ICARDA-supplied breeding material. 7.7.2 Conventional Breeding Efforts

7.7.2.1 Breeding MethodsThe breeding methods used in chickpea are not different from other self-pollinated food legumes. In the early phase of chickpea breeding, most varieties were developed through selection from the landraces collected in the country or through germplasm introduction, evaluation, and selection. Later, the emphasis gradually shifted to hybridization for increasing genetic variation in the breeding materials. Most recent varieties have been developed through hybridization. Single, three-way, double, or multiple crosses are used, depending on the number of traits to be combined and their spread in the parents. It is well established that selection for yield in early segregating generations is not effective in chickpea because of its indeterminate growth habit. Thus, many scientists prefer to select crosses rather than plants within crosses in early segregating generations (F2 and F3). The pedigree method, earlier used at many institutes, including ICRISAT, is not practiced widely in its original form, because it is cumbersome and only a limited number of crosses can be handled by this method. The bulk method, variously modified, is now the most common selection method used after hybridization in chickpea. Single-seed descent (SSD) method, a modification of bulk method, is widely used for development of recombinant inbred lines (RILs) for genome mapping studies. Backcross method is commonly used to incorporate one or few traits from a germplasm line, sometimes a wild species, to a well-adopted variety. A population improvement method that involves intercrossing of selected plants in F2s or F3s has been suggested for legumes for enhancing the chances of recombination in segregating generations (Muehlbauer et al., 1988). van Rheenen et al. (1991) proposed a method called polygon breeding, whereby segregating populations and selections are shared and exchanged between breeders. This method has been used by ICRISAT in collaboration with some state agricultural universities in India. Attempts have been made to define chickpea ideotypes for different growing conditions. In the drought-prone environments, traits that help plants to escape or tolerate drought should be considered in the ideotype. Saxena and Johansen (1990) suggested that an ideotype for drought-stress environments should have early maturity, a deep root system, and a smaller leaf size. In the Mediterranean climate, the crop experiences a cool and wet winter followed by rapid warming in spring, leading to terminal drought. Thus, the ideotype for the Mediterranean environments should


Figure 7.1 (See color insert following page 178) Breeding chickpea for early maturity at ICRISAT. Short-duration varieties have helped adaptation of chickpea in tropical environments. include early flowering and tolerance to cold during flowering (Sedgley et al., 1990). It has also been suggested that a compact plant type with erect growth habit and short internodes could help resist excessive growth in high input conditions (Dahiya and Lather, 1990). A spontaneous mutant with short internodes and compact growth habit, E100YM, has been identified (Dahiya et al., 1984) and used in ideotype breeding. Promising progenies with compact growth habit have been obtained, which can be grown at high plant density (Lather, 2000).

7.7.2.2 Breeding for Tolerance to Abiotic Stresses and Widening of

AdaptationDrought is the most important constraint to yield in chickpea accounting for 40 to 50% yield reduction globally. Four approaches are being pursued: (1) high root mass, (2) smaller leaf area, (3) osmoticum adjustment, and (4) early-maturing short-duration varieties. Lines with a greater degree of drought tolerance have been developed by combining large root traits of ICC 4958 with fewer pinnules trait of ICC 5680 (Saxena, 2003). A recent screening of the mini-core collection has identified several other lines with large root traits (Krishnamurthy et al., 2003). Genotypic variation for osmotic adjustment has been observed in chickpea (Morgan et al., 1991; Leport et al., 1999; Abbo et al., 2002b). The heritability of osmotic adjustment has been found to be low (h2 =0.20 to 0.33), indicating that gains from selection for increased osmotic adjustment are likely to be small (Abbo et al., 2002b). ICRISAT has placed a high emphasis on development of short-duration varieties (Figure 7.1), as these can escape terminal drought. The first

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breakthrough was the development of an extra-short-duration kabuli variety, ICCV 2, from a multiple cross involving five parents (2 kabuli and 3 desi), which has helped the expansion of kabuli chickpea cultivation to tropical environments. Now ICRISAT has developed super-early chickpea lines (ICCV 96029, 96030) in desi chickpea, and these are being used extensively in crossing programs by NARS in India and Canada for development of early varieties. Singh (1990) listed several advantages of winter chickpea in comparison to spring chickpea, including higher germination rate, less incidence of fusarium wilt, increased nitrogen fixation, better utilization of available moisture, mechanization of harvesting due to increased crop height, and increased protein and grain yield per hectare. The effects of low-temperature stress and its implications for chickpea improvement were reviewed recently by Croser et al. (2003b). Most chickpea cultivars are susceptible to chilling temperature at flowering (Croser et al., 2003b). The germplasm line ILC 8262, the mutant ILC 8617, and the breeding line FLIP87–82C were the best sources of cold tolerance in cultigen, with a consistent score of 3 (on a 1-to-9 scale) over years and locations (Singh et al., 1990, 1995). A number of varieties combining cold tolerance and resistance to ascochyta blight have been released in several countries from the breeding material supplied by ICARDA. A pollen selection method, as described by Clarke et al. (2004), has also been successfully used to transfer cold tolerance from ICCV 88516 (CTS 60543) to the popular varieties Amethyst and Tyson in Australia (Clarke and Siddique, 2004).Legumes, in general, are sensitive to salinity, and within legumes, chickpea, faba bean, and field pea are more sensitive than other grain legumes. Saline soils are very common in West and Central Asia and Australia, where chickpea is widely grown. Soil salinity adversely affects germination, resulting in poor plant stand. Only salt tolerant cultivars can be grown successfully in soils having ECe higher than 4.0 dS/m. The levels of salinity tolerance identified in chickpea are low to moderate. Several tolerant sources have been identified in India and Pakistan (Singh and Singh, 1984; Dua and Sharma, 1995; Kathiria et al., 1997). The salt-tolerant lines CSG 88101and CSG 8927 identified by Dua and Sharma (1995) had lower Na+ in root than the sensitive genotypes. A salinity tolerant desi chickpea variety Karnal Chana 1 (CSG 8963) has been released in India for salt-affected soils of northwestern parts. 7.7.2.3 Breeding for Resistance to Biotic Stresses

7.7.2.3.1 Diseases Ascochyta blight (AB), caused by Ascochyta rabiei (Pass.) Labr. and represented by several pathotypes, is a highly devastating foliar disease of chickpea in West and Central Asia, North Africa, North America, and Australia. The problem of fungal variability and the existence of races appear to be complex. A standard set of well-characterized genotypes, a common inoculation technique, and a well-defined disease-rating methodology should be used by workers who wish to determine the extent and distribution of variability in A. rabiei in different geographic regions (Haware, 1998). Considerable efforts have been made on identification of chickpea genetic resources resistant to AB and breeding for AB resistance. Multilocation evaluation of chickpea


germplasm indicated that kabuli germplasm generally shows higher resistance to AB than desi germplasm (Reddy et al., 1992; Haware et al., 1995). Extensive germplasm screening has identified many resistant desi and kabuli lines (Reddy and Singh, 1984; Singh et al., 1984b). Using the breeding material supplied by ICARDA and ICRISAT, more than 100 AB tolerant varieties have been released in 25 countries. The national breeding programs in the U.S., Canada, Australia, Pakistan, India, Europe, and Western Asia have also made good progress in development of AB-resistant varieties. Fusarium wilt (FW) is the most important root disease of chickpea. It has been reported from almost all the chickpea-growing regions of the world. Seven races of FW have been reported worldwide (Phillips, 1988; Jimenez-Diazet, et al., 1989). Effective field, greenhouse, and laboratory procedures for resistance screening have been developed (Nene et al., 1981) (Figure 7.2), and good progress has been made in identifying sources of resistance (Haware et al., 1990; van Rheenen et al., 1992). A number of varieties with absolute resistance to FW are available in many countries. In most cases, the resistance to FW has been stable. Some of the varieties released in India about two decades ago, such as JG 315 and JG 74, still maintain a high level of resistance.Botrytis gray mold (BGM), caused by the necrotrophic fungus Botrytis cinerea Pres., is an important foliar disease of chickpea in northern India, Nepal, Bangladesh, and Pakistan (Haware and McDonald, 1992) and has been reported from more than 15 countries (Nene et al., 1996), including Australia (Corbin, 1975) and North America (Kharbanda and Bernier, 1979). High levels of resistance have not been found in the cultivated species (Haware and Nene, 1982; Haware and McDonald, 1993). Some accessions with erect plant type, such as ICCL 87322 and ICCV 88510, were found to be less affected by the disease (Haware and McDonald, 1993).

Figure 7.2 A Fusarium wilt screening nursery at ICRISAT.

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Three types of root rot diseases (dry root rot, collar rot, and phytophthora root rot) are known to affect chickpea. High soil moisture, the presence of undecomposed organic matter on the soil surface, low soil pH, and a high temperature favor collar rot. There has been limited research toward the identification of races of dry root rot, even though the existence of different races has been implicated (Than et al., 1991). Pundir et al. (1988) provided a list of 47 tolerant accessions available in the genebank of ICRISAT. A few germplasm lines and cultivars have been identified to have low to moderate levels of resistance. These include the cultivars SAKI 9516 (Dua et al., 2001), breeding lines RSG 130, 132, and 191 (Chitale et al., 1990), and germplasm accessions ICC 1696, ICC 4709, and ICC 14391 (Singh, personal communication, 2003). The breeding efforts have led to release of a number of moderately resistant varieties, such as ICCC 37, ICCV 10, JG 130, and WCG 1 (Dua et al., 2001). Phytophthora root rot, caused by Phytophthora

medicaginis Hansen, is the major disease of chickpea in northern New South Wales and Queensland (Knights et al., 2003). Genotypic differences in resistance to Phytophthora root rot have been identified (Brinsmead et al., 1985; Dale and Irwin, 1991), and cultivars less susceptible to the disease, such as Jimbour, have been developed. The levels of resistance available in the cultivated species are low as compared with the wild species (Knights et al., 2003). 7.7.2.3.2 Root Nematodes and Insect Pests The major nematodes known to affect chickpea are root knot nematodes (Meloidogynespp.), cyst-forming nematodes (Heterodera spp.), and lesion nematodes (Prathylenchusspp.). None of the 8000 accessions of cultivated chickpea (7000 kabuli type and 1000

desi type) screened at ICARDA against cyst nematodes was found to have even a moderate level of resistance (Di Vito et al., 1988). However, good sources of resistance have been identified in the wild species (Robertson et al., 1995). Pod borer (Helicoverpa armigera Hubner) is the most important insect pest of chickpea worldwide. Several techniques are available for screening of chickpea genotypes for resistance to pod borer (Sharma et al., 2003). Only low to moderate levels of resistance for pod borer have so far been identified in the cultivated and wild Cicerspecies (Lateef and Pimbert, 1990; Sharma et al., 2002). Unavailability of a high level of resistance to pod borer in the cultivated and cross-compatible wild species has been the major limitation in developing pod borer resistant varieties. Limited progress has been made in breeding for resistance to leaf miner (Malhotra et al., 1996; Robertson et al., 1995). Seed beetle or bruchid (Callosobruchus spp.) is the most important storage pest of chickpea. No source of resistance to bruchid has been identified in the cultivated species. Some of the accessions of wild species showed no damage by this insect (Robertson et al., 1996). It is yet to be established whether the resistance is due to seed morphology or chemical properties. 7.7.2.4 Introgression of Economically Valuable Traits through Wide

CrossesInterspecific hybridization has played an important role in genetic enhancement of many crop species by facilitating transfer of useful traits to the cultivated species from wild,


weedy forms. However, the progress in chickpea is still in its infancy (Croser et al., 2003a). Although the wild annual Cicer species have been extensively evaluated, their exploitation in chickpea breeding programs has not yet been widely realized. No perennial species has yet been successfully hybridized with the cultivated species. The exploitation of the wild Cicer species in breeding programs has been limited to cyst nematode (Heterodera ciceri Vovlas, Greco & Di Vito) resistance (Di Vito et al., 1996; Singh et al., 1996; Erskine et al., 2001; Malhotra et al., 2002). Selections derived from the interspecific hybrid of C. arietinum×C. echinospermum and possessing resistance to root lesion nematode, enhanced phytophthora resistance, and moderate resistance to ascochyta blight are also being considered for release in Australia (Knights et al., 2002). In addition to the genes for resistance and tolerance, heterosis for yield has been observed in populations from crosses between C. arietinum, C. reticulatum, and C.

echinospermum (Jaiswal et al., 1987; Singh and Ocampo, 1993, 1997). While, most introgressions have involved the two crosscompatible species, C. reticulatum or C.

echinospermum, Singh et al. (1994), and Verma et al. (1995) are the only ones reporting F2 recombinants with a very large number of secondary branches, high pod number, and yield from a cross between C. arietinum and C. judaicum, or C. pinnatifidum, or C.

bijugum. Some progress has also been made in producing early maturing segregants, following interspecific hybridization. Transgressive segregants for days to flowering have been isolated from the interspecific cross C. arietinum×C reticulatum (Singh et al., 1984a).An interesting perspective has been brought forth by Abbo et al. (2002a), who recently presented evidence indicating that some wild Cicer relatives have a vernalization requirement. This may be useful in certain environments to avoid early flowering and consequent frost damage, etc.; however, it also can be an unfavorable trait in other environments. Introgression of genes from the wild species back into chickpea may result in reintroduction of these vernalization-sensitive alleles into the cultivated species and should be considered with caution in any hybridization program. 7.7.2.5 Mutation BreedingMost studies on polymorphism of molecular markers in chickpea indicate presence of limited genetic variability in the cultivated species, and this has forced researchers to use interspecific crosses for genome mapping (Gaur and Slinkard, 1990a, 1990b; Kazan et al., 1993; Simon and Muehlbauer, 1997; Winter et al., 1999, 2000). Kenneth J.Frey (Iowa State University, Ames, IA) called chickpea “a recalcitrant crop species,” as it has not been very amenable to genetic improvement in spite of extensive breeding efforts during the past three decades (van Rheenen et al., 1993). It was suggested that mutation breeding and interspecific hybridization should be used for increasing genetic variability and yield advancement of chickpea (van Rheenen et al., 1993). A large variability is seen in chickpea germplasm for morphological traits, but it could be a reflection of the expression of a limited number of mutant genes, as a single mutant gene may cause marked changes in the appearance of the plant (Gaur and Gour, 2003). Though a variety of physical and chemical mutagens have been used, the most commonly used mutagen is -rays, with doses ranging from 10 to 40 kR. There are reports

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suggesting that kabuli chickpeas are more sensitive to mutagens than the desi type (Khanna and Meharchandani, 1981; Kharkwal, 1998). At least two institutes, the Nuclear Institute of Agriculture and Biology (NIAB), Faisalabad, Pakistan and the Indian Agriculture Research Institute (IARI), New Delhi, India, have had strong programs on mutation breeding for incorporating disease resistance for chickpea improvement. The first of the three desi-type Ascochyta blight-resistant varieties, CM 72 from the NIAB program, was released in 1983, which was then followed by CM 88 in 1994, and CM 98 in 1998 (Haq et al., 1999). A kabuli variety, CM 2000, from this program was released in 2000. In India, at least six chickpea varieties have been developed through mutation breeding. Of these, three were developed by IARI. The mutation breeding program of IARI has had a major focus on resistance to diseases. The varieties, Pusa 408 (Ajay) and Pusa 413 (Atul), were moderately resistant to AB, whereas Pusa 417 (Girnar) was resistant to wilt and moderately resistant to stunt and root rots (Kharkwal et al., 1988; Micke, 1988; Dua et al., 2001). Three other chickpea mutants, RS 11, RSG 2 (Kiran), and WCG 2 (Surya), were developed by agricultural universities (Micke, 1988; Dua et al., 2001). Mutation breeding has also helped to improve the nutritional quality of chickpea. The Bangladesh Institute of Nuclear Agriculture, Mymensingh, obtained a high-yielding and high-protein mutant of chickpea cultivar Faridpur-1 through -rays induced mutation. The mutant had 20% higher yield and 20% higher protein than the parental cultivar Faridpur-7. The mutant was released as a cultivar by the name “Hyprosola” in 1981 by the National Seed Board of Bangladesh (Oram et al., 1987). These examples clearly indicate that mutation breeding has played important role in chickpea improvement. In addition to the release of 11 mutants as cultivars, several mutants have been used as parents in breeding programs. 7.7.2.6 Development of Hybrids Using Cytoplasmic Male Sterility (CMS)

SystemMale sterility has been reported in chickpea (Chaudhary et al., 1970; Sethi, 1979). Genetic studies indicated that the male sterility was under the control of a monogenic recessive gene (Chaudhary et al., 1970; Reddy and Reddy, 1997). It has also been possible to induce male sterility (up to 80% pollen sterility) through gametocide (Mathur and Lal, 1999). Utilization of male sterility for hybrid seed production requires an efficient mechanism for pollen dispersal by the male parent and reception of pollen by the stigma of the male sterile line. This is the biggest hurdle in chickpea, where cross-pollination has been reported to be less than 1% (Tayyar et al., 1995) due to the cleistogamous nature of the flower. Thus, the availability of CMS and fertility restorer systems will not be enough in chickpea for production of hybrid seed. A significant change in floral morphology is needed in the A, B, and R lines. It basically requires open flowers in which stigma and anthers are not enclosed by the petals. A male sterile open flower mutant has been reported in chickpea (Pundir and Reddy, 1988). It may be used as female parent. A similar, but male fertile, mutant is needed for use as male parent. A fertile outwardly curved-wing mutant has been reported (Gaur and Gour, 2003), but the keel petal still encloses the anthers in this mutant.


The commercial cultivation of hybrid chickpea may still be far from reality, even if we have the appropriate CMS system and the required changes in floral morphology. It will be difficult to produce a large quantity of hybrid seed as each pod produces only one or two seeds, and seed-to-seed multiplication is notoriously low in chickpea and other cool season grain legumes, compared to cereals and canola. Most chickpea cultivation (95%) is in developing countries where the majority of farmers cannot afford expensive hybrid seed every year. As there is no immediate hope of utilization of a male sterility system for development of commercial hybrids in chickpea, apomixis can be explored as an alternative system for exploitation of heterosis in this important food legume. So far, however, there is no report detailing the occurrence of apomixis in chickpea. 7.7.3 Genetic TransformationThe advancement in recombinant DNA technology and availability of efficient transformation and regeneration systems in plants have made it possible to transfer genes from any organism to plants with optimized expression. Targeted transfer of genes from the wild Cicer species into the cultivated species would represent a very elegant application of transformation technology. The progress in molecular mapping and, to some extent, chickpea transformation, now considered a routine procedure in chickpea, has brought the application of this type of technology much closer to reality in chickpea (Fontana et al., 1993; Hamblin et al., 1998; Chakrabarty et al., 2000; Krishnamurthy et al., 2000; Sharma and Ortiz, 2000; Jaiwal et al., 2001; Senthil et al., 2004). The applicability of this technology will, however, depend on the identification of key genes, the number of genes conferring a particular character, and public acceptance of cultivars resulting from transformation technology. In chickpea, transgenic technology is being exploited primarily for insectpests and disease resistance, drought tolerance, and quality enhancement. An efficient and reproducible tissue culture regeneration system is a prerequisite for development of transgenics. Many earlier studies on chickpea tissue culture encountered problems in rooting or establishment of plants in the soil. One option tried was to graft the in vitro germinated shoot on scions of pre-germinated seedling (Krishnamurthy et al., 2000), although such a tedious method would be inefficient for routine use. Novel rooting systems that give a rooting frequency of 90% or more have been reported. These involve either placing elongated shoots on a filter paper bridge immersed in liquid rooting medium (Jayanand et al., 2003) or placing nodal segments in an inverse polarity in a specially formulated tissue culture medium (Fratini and Ruiz, 2003). Several studies have been conducted on transformation of chickpea during the past decade. Reports are now available on successful transformation of chickpea and regeneration of transformed plants. The most commonly used method has been the

Agrobacterium-mediated transformation using embryo axes as explants (Fontana et al., 1993; Kar et al., 1996; Chakrabarty et al., 2000; Krishnamurthy et al., 2000; Chandra and Pental, 2003). However, bombardment with accelerated tungsten particles has also been successfully used (Hussain et al., 1997; Kar et al., 1997; Tewari-Singh et al., 2004). Except Kar et al. (1997), all these studies used only marker genes like uidA and npt II for transformation. Sommers et al. (2003) are of the opinion that, in addition to the limited availability of an efficient transformation protocol, a rapid and reliable selection strategy

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has also impeded the uptake of this technology in chickpea improvement programs. An efficient and reliable nonantibiotic selection strategy using the phosphinothricin-acetyltransferase and aspartate kinase genes, has been recently developed for the production of transgenic chickpea (Tewari-Singh et al., 2004). One of the areas, where traditional chickpea breeding methods have yet to make progress is the development of cultivars resistant to the devastating pest helicoverpa pod borer. It is simply because sources of a high level of resistance are not available in the cultivated or cross-compatible wild species. Thus, development of transgenic chickpea seems to be the only hope to host-plant resistance to this insect-pest. There is a report (Kar et al., 1997) on development of transgenic chickpea plants for BtCry1A(C) gene derived from the bacterium Bacillus thuringiensis (Bt). The transformation method used was particle bombardment of embryo axis (devoid of root and shoot meristems) from mature seeds. The transformation was confirmed through molecular analysis. Insect feeding assay indicated inhibition of development of feeding larvae. There is no further report available on field testing of these transgenics. ICRISAT has produced transgenics for resistance to pod borer by using BtCry1Ab and SbTi (soybean trypsin inhibitor) genes. The molecular characterization and insect bioassays are currently ongoing (Sharma, personal communication, 2003). New constructs with combinations of different Cry genes are being developed in collaboration with CIRAD (Centre de Coopération Internationale en Recherche Agronomique pour le Développement) France, besides the artificial synthesis of some of these genes for optimal expressions in plants (Lavanya et al., 2003). Research at the Scottish Crops Research Institute, U.K., has shown that immature raspberry fruit contains a polygalacturonase inhibitory protein (PGIP) effective against endopolygalactoseuronases produced constitutively by Botrytis cinerea (Johnston et al., 1993). Polygalactuseuronases are the key enzymes in the invasion of plant tissues by many facultative fungal pathogens. At ICRISAT, PGIP gene and other antifungal genes, such as chitinases and glucanases, are being introduced into chickpea for resistance to fungal diseases (Sharma and Ortiz, 2000). Efforts are also being made to identify and clone tissue-specific promoters for more controlled expression of these potential transgenes. A project funded under the Indo-Swiss Collaboration in Biotechnology (ISCB) is aimed at developing transgenic chickpea for tolerance to drought and low-temperature stresses by using genes with regulatory functions, such as drought responsive elements (DREs), and osmoregulation, such as codA and P5CSF (Sharma, personal communication, 2003). Research in progress in this area is expected to improve chickpea for the traits that are difficult to improve through traditional breeding methods. The applicability of this technology will, however, depend on the identification of key genes, the number of genes conferring a particular character, and public acceptance of cultivars resulting from transformation technology. In addition, Sagare and Krishnamurthy (1991) have proposed fusion of protoplasts as an alternative means of enabling interspecific hybridization between widely related Cicerspecies. This technique is currently limited by the absence of protocols for the regeneration of whole plants from single cells or protoplasts in any of the Cicer species. Recent progress toward the development of a microspore culture system in Cicer (Croser et al., 1999; Lülsdorf et al., 2001; Croser, 2002; Croser and Lülsdorf, 2004) resulting in


embryogenesis from single-celled microspores in liquid culture may assist in the future development of this technique. 7.7.4 Molecular Marker-Assisted SelectionThe underlying principle of molecular marker-assisted selection is that if a gene(s) is linked to an easily identifiable genetic marker, then it may be more efficient to select in a breeding program. This effectiveness, however, will depend on the strength of linkage of the marker to the locus controlling the character of interest. Such an approach can be the most effective means of enhancing the transfer of a desired gene(s) by a backcross program, reducing the linkage drag resulting from a wide cross, and for pyramiding genes and alleles of various combinations. The successful application of marker technologies, thus, requires (1) a dense saturated intraspecific map linking traits of agronomic importance to highly polymorphic, co-dominant markers in sufficiently close proximity to allow marker-assisted selection in offspring derived from intraspecific crosses, and (2) tailoring of markers and reactions in combination with high-thoroughput screening techniques to speed up and facilitate the application of marker technology. The history of developing and identifying molecular markers for marker-assisted selection in chickpea, understandably reflects a mirror image of the increasing understanding of genetic linkage maps in the crop. The first of such markers was not identified until 1997 (Mayer et al., 1997), however, recent progress has resulted in many markers being identified (Table 7.4). Indeed, the developments in chickpea have lagged considerably behind those in cereals (Gupta et al., 1999). Other than the molecular markers identified for double podding trait (Cho et al., 2002; Rajesh et al., 2002a), leaf traits, and erect growth habit (Banerjee et al., 2001), only genes conferring resistance to fusarium wilt and ascochyta blight have been tagged. Most of the populations utilized in genetagging experiments have been derived from interspecific crosses with the wild annual species C. reticulatum and, to a limited extent, C. echinospermum, as well as some intraspecific crosses within the cultivated species. Among the various types of molecular markers that are available, the STMS markers are most amenable for multiplex PCR and electrophoresis, as shown for more than 600 STMS markers in soybean (Cregan et al., 1999; Narvel et al., 2000). In chickpea, the number of STMS markers is still too low for such applications. Marker development is in progress at the various research institutions working on chickpea, however, there is no application of a robust

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Table 7.4 Potentially Useful Markers for Marker-Assisted Selection in Chickpea ImprovementPublication Marker RemarksMayer et al., 1997 RAPD marker UBC-170550 7 cM from fusarium wilt resistance to race 1

Ratnaparkhe et al., 1998b RAPD marker UBC-855500 Linked in repulsion, 5.2 cM from fusarium wilt resistance to race 4 Ratnaparkhe, et al.,1998b ISSR marker UBC-8251200 5.0 cM from fusarium wilt resistance to race 4 Tullu et al., 1998 RAPD marker CS-27700 9 cM from fusarium wilt resistance to race 1 Tullu et al., 1999 RAPD marker CS-27700 Fusarium wilt resistance to race 4 Santra et al., 2000 2 RAPD marker 10.9 cM apart and flanking major QTL-1 for ascochyta blight resistance 1 ISSR and 1 isozyme 5.9 cM apart and flanking major QTL-2 for ascochyta blight resistance Banerjee et al., 2001 RAPD markers Linked to QTL for leaf length, leaf width, and erect plant habit Cho et al., 2002 PCR marker Tr44 7.8 cM from gene for double podding trait QTL marker Ta130s Explaining 31% of the variation for seed number per plant Rajesh et al., 2002a STMS marker T-80 4.84 cM from gene for double podding trait Benko-lseppon et al., 2003 SCAR marker R-2609–1 2.0 cM from fusarium wilt resistance to race 4 Collard et al., 2003b QTL marker CS44a1150 Significant QTL for seedling resistance to ascochyta blight Flandez-Galvez et al., 2003b 6 QTL markers For ascochyta blight resistance Idnani and Gaur, 2003 Isozyme marker Amy 6.2 cM from leaf necrosis gene (nec)Millan et al., 2003 19 RAPD, 1 ISSR markers Distributed in an 18.8 cM region, and resistance to ascochyta blight, QTL marker marker OPAC/1200 present most frequently Rakshit et al., 2003 DAF marker OPS06–1 Significant QTL for ascochyta blight resistance


Rubio et al., 2003 RAPD marker OPJ20600 6 cM from fusarium wilt resitance to race 0 Sharma et al., 2004 STMS markers TA96, TA27 0.6 cM from fusarium wilt resistance to race 3 molecular marker system for selection within a large breeding program. We anticipate that this technology is not far away, and it holds great potential for targeted introgression of genes from wild Cicer into chickpea.

7.8 SUMMARYThe rich diversity for various biotic and abiotic stresses present in the annual and perennial wild related species has to be exploited for genetic enhancement of chickpea. Wide hybridization is currently limited by reproductive barriers and simultaneous transfer of undesirable characters. The perennial species, in particular, are poorly evaluated and in many ways an unknown quantity. There is a clear need to develop methods for the maintenance and creation of ex situ collections of perennial species to enable their evaluation for important agronomic traits. An efficient embryo rescue protocol to enable crosses between chickpea and the more distantly related tertiary gene pool Cicer species will be crucial to the utilization of traits from these wild relatives. Recent advances in biotechnology techniques, including embryo rescue for interspecific hybrids, linkage mapping, marker-assisted breeding, and transformation, hold much promise for successful introgression of genes from wild annual species into cultivated chickpea in the future. The development and commercialization of genetically transformed chickpea, however, should be approached with caution, keeping in mind their possible rejection by consumers. There is little doubt that wild related Cicer species will play an important role in the future improvement of chickpea varieties. The task at hand is to identify the novel genes and find the best strategy to utilize them in a breeding program. The challenges facing the utilization of Cicer genetic resources are global and call for a wider collaborative research initiative for maximizing resource management and gains.

ACKNOWLEDGMENTSThe authors would like to thank the following funding agencies for supporting chickpea research: Saskatchewan Pulse Growers Association (SPGA), Grains Research and Development Corporation (GRDC), Centre for Legumes in Mediterranean Agriculture (CLIMA), Brandon University Research Committee (BURC), Agriculture Development Fund (ADF)-Saskatchewan, and various donors to ICRISAT’s chickpea research.

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REFERENCESAbbo, S., Berger, J., and Turner, C., Evolution of cultivated chickpea: four bottlenecks limit diversity and constrain adaptation, Functional Plant Biol, 30, 1081, 2003. Abbo, S., Lev-Yadun S., and Galwey, N., Vernalization response of wild chickpea, New

Phytologist, 154, 695, 2002a. Abbo, S. et al., Detection of ribosomal DNA sites in lentil and chickpea by fluorescent in situhybridization, Genome, 37, 713, 1994. Abbo, S. et al., Breeding for osmotic adjustment in chickpea (Cicer arietinum L.). In Proceedings

of the 12th Australian Plant Breeding Conference, McComb, J.A., Ed., Australian Plant Breeding Association, Perth, Australia, 463, 2002b. Ahmad, F., Interspecific hybridization and genetic relationships among the annual Cicer L. species, Ph.D. Thesis, University of Saskatchewan, Saskatoon, 1988. Ahmad, F., The chromosomal architecture of Cicer anatolicum Alef., a wild perennial relative of chickpea, Cytologia, 54, 753, 1989. Ahmad, F., Random amplified polymorphic DNA (RAPD) analysis reveals genetic relationships among the annual Cicer species, Theor. Appl. Genet., 98, 657, 1999. Ahmad, F., A comparative study of chromosome morphology among the nine annual species of Cicer L., Cytobios, 101, 37, 2000. Ahmad, F. and Chen, Q., Meiosis in Cicer L. species: The relationship between chiasma frequency and genome length, Cytologia, 65, 161, 2000. Ahmad, F. and Hymowitz, T., The fine structure of chickpea (Cicer arietinum L.) chromosomes as revealed by pachytene analysis, Theoret. Appl. Genet., 86, 637, 1993. Ahmad, F. and Kollipara, K.P., unpublished data, 2004. Ahmad, F. and Slinkard, A.E., Genetic relationships in the genus Cicer L. as revealed by polyacrylamide gel electrophoresis of seed storage proteins, Theor. Appl. Genet., 84, 688, 1992. Ahmad, F. and Slinkard, A.E., Limitations to bridge-species facilitated alien gene transfers in chickpea: prefertilization events, J. Genet. Breed., 57, 69, 2003. Ahmad, F. and Slinkard, A.E., The extent of embryo and endosperm growth following interspecific hybridization between Cicer arietinum L. and related annual wild species, Genetic Resources

Crop Evol, 51, 765, 2004. Ahmad, F., Gaur, P.M., and Slinkard, A.E., Isozyme polymorphism and phylogenetic interpretations in the genus Cicer L., Theor. Appl Genet., 83, 620, 1992. Ahmad, F., Slinkard, A.E., and Scoles, G.J., The cytogenetic relationship between Cicer judaicumBoiss. and Cicer chorassanicum (Bge.) M. Pop., Genome, 29, 883, 1987. Ahmad, F., Slinkard, A.E., and Scoles, G.J., Investigations into the barrier(s) to interspecific hybridization between Cicer arietinum L. and eight other annual Cicer species, Plant Breed.,100, 193, 1988. Arora, P.P., Genetic variability and its relevance in chickpea improvement, International Chickpea

Newslt., 25, 9, 1991. Arumuganathan, K. and Earle, E.D., Nuclear DNA content of some important plant species, Plant

Mol. Biol Rept., 9, 208, 1991. Badami, P.S., Mallikarjuna, N., and Moss, J.P., Interspecific hybridization between Cicer arietinumand C. pinnatifidum, Plant Breed., 116, 393, 1997. Bahl, P.N., Cytology of chickpea. In The Chickpea, Saxena, M.C., and Singh, K.B., Eds., CAB International Publications, Oxon, U.K., 83, 1987.Banerjee, H., Pai, R.A., and Sharma, R.P., Restriction fragment length polymorphism and random amplified polymorphic DNA analysis of chickpea accessions, Biologia Plantarum, 42, 197, 1999.


Banerjee, H. et al., Use of random amplified polymorphic DNA markers for mapping the chickpea genome, Biologia Plantarum, 44, 195, 2001. Bassiri, A., Ahmad, F., and Slinkard, A.E., Pollen grain germination and pollen tube growth following in vivo and in vitro self and interspecific pollinations in annual Cicer species, Euphytica, 36, 667, 1987. Benko-Iseppon, A.M. et al., Molecular markers closely linked to fusarium resistance genes in chickpea show significant alignments to pathogenesis-related genes located on Arabidopsischromosomes 1 and 5, Theor. Appl. Genet., 107, 379, 2003. Berger, J., Abbo, S., and Turner, N.C., Ecogeography of annual wild Cicer species: The parlous state of the world collection, Crop Sci., 43, 1076, 2003. Brinsmead, R.B. et al., Resistance in chickpea to Phytophthora megaperma f. sp. Medicagimis,

Plant Disease, 69, 504, 1985. Chakrabarty, R., Singh, D.T., and Garg, G.K., Transformation studies on chickpea embryo axis, J.

Plant Biochem. Biotechnol., 9, 107, 2000. Chandel, K.P.S., A note on the occurrence of wild Cicer microphyllum Benth. and its nutrient status, International Chickpea Newslt., 10, 4, 1984. Chandra, A. and Pental, D., Regeneration and genetic transformation of grain legumes: An overview, Current Sci., 84, 381, 2003. Chaudhary, M.S., Chandra, S., and Tomar, Y.S., Genetical studies on a variant form in Bengal gram (Cicer arietinum L.), Indian J. Hered., 2, 133, 1970. Chitale, K., Tyagi, R.N.S., and Singh, R.D., Reaction of chickpea cultivars to collar rot disease induced by Sclerotium rolfsii, Indian J. Mycology Plant Pathol., 20, 262, 1990. Cho, S. et al., Mapping genes for double podding and other morphological traits in chickpea, Euphytica, 128, 285, 2002. Choumane, W. et al., Conservation and variability of sequence tagged microsatellite sites (STMSs) from chickpea (Cicer arietinum L.) within the genus Cicer, Theor. Appl. Genet., 101, 269, 2000. Clarke, H.J. and Siddique, K.H.M., Response of chickpea genotypes to chilling tolerance during reproductive development, Field Crops Res., 90, 23, 2004. Clarke, H.J., Khan, T.N., and Siddique, K.H.M., Pollen selection for chilling tolerance at hybridisation leads to improved chickpea cultivars, Euphytica, (in press), 2004. Collard, B.C.Y., Pang, E.C.K., and Taylor, P.W.J., Selection of wild Cicer accessions for the generation of mapping populations segregating for resistance to ascochyta blight, Euphytica,130, 1, 2003a. Collard, B.C.Y. et al., Prospecting for sources of resistance to ascochyta blight in wild Cicerspecies, Australian J. Plant Pathol., 30, 271, 2001. Collard, B.C.Y. et al., Preliminary investigation of QTLs associated with seedling resistance to ascochyta blight from Cicer echinospermum, a wild relative of chickpea, Theor. Appl. Genet.,107, 719, 2003b. Corbin, E.J., Present status of chickpea research in Australia. In Proceedings of the International

Workshop on Grain Legumes, ICRISAT, Patancheru, India, 97, 1975. Cregan, P.B. et al., An integrated linkage map of the soybean genome, Crop Sci., 39, 211, 1999. Croser, J.S., Haploid and zygotic embryo culture of Cicer, Ph.D. Thesis, University of Melbourne, Australia, 2002. Croser, J.S. and Lülsdorf, M., Progress toward a doubled haploid protocol for pea, chickpea and lentil. Proceedings of the European Grain Legumes Conference, Dijon, France, June 7-11, 189, 2004.Croser, J.S., Lülsdorf, M., and Vandenberg, A., Isolated microspore culture for the development of doubled haploids in chickpea (Cicer arietinum L.). In Proceedings National Pea Improvement

Association BiAnnual Conference, Calgary, AB, 1999. Croser, J.S. et al., Utilisation of wild Cicer in chickpea improvement—progress, constraints, and prospects, Australian J. Agricultural Res., 54, 429, 2003a.

Chickpea 257

Croser, J.S. et al., Low-temperature stress: Implications for chickpea (Cicer arietinum L.) improvement, Critical Reviews in Plant Sciences, 22, 185, 2003b. Cubero, J.I., Morphology. In The Chickpea, Saxena, M.C. and Singh, K.B., Eds., CAB International, Cambridge, U.K., 35, 1987. Cubero, J.I. and Ocampo, B., personal communication, 2004. Dahiya, B.S. and Lather, V.S., A breakthrough in chickpea yields—Paper 1, International

Chickpea Newslt., 22, 6, 1990. Dahiya, B.S. et al., Useful spontaneous mutants in chickpea, International Chickpea Newslt., 11, 5, 1984.Dale, M.L. and Irwin, J.A.G., Glasshouse and field screening of chickpea cultivars for resistance to Phytophthora megasperma f. sp. medicaginis, Australian J. Exptl. Agril., 31, 663, 1991. De Leonardis, W., Fichera, G., and Zizza, A., Pollen and seed morphology of Cicer arietinum L. cultivars and relationships with C. reticulatum Ladiz. and C. echinospermum P.H. Davis, Plant

Genetic Resources Newslt., 105, 29, 1996. Di Vito, M. et al., Response of chickpea germplasm lines to Heterodera ciceri attack, Nematologia

Mediterranea, 16, 17, 1988. Di Vito, M. et al., Sources of resistance to cyst nematode in cultivated and wild Cicer species, Genetic Resources Crop Evol., 43, 103, 1996. Dua, R.P. and Sharma, P.C., Salinity tolerance of kabuli and desi chickpea genotypes, Int.

Chickpea and Pigeonpea Newsl., 2, 19, 1995. Dua, R.P., Chaturvedi, S.K., and Shiv Sewak, Reference Varieties of Chickpea for IPR Regime,Indian Institute of Pulses Research, Kanpur, 2001. Erskine, W. et al., Utilisation of grain legume diversity. In Plant Genetic Resources of Legumes in

the Mediterranean, Maxted, N. and Bennett, S.J., Eds., Kluwer Academic Publishers, Dordrecht, The Netherlands, 311, 2001. Flandez-Galvez, H. et al., An intraspecific linkage map of the chickpea (Cicer arietinum L.) genome based on sequence tagged microsatellite site and resistance gene analog markers, Theor.

Appl. Genet., 106, 1447, 2003a. Flandez-Galvez, H. et al., QTL analysis for ascochyta blight resistance in an intraspecific population of chickpea (Cicer arietinum L.), Theor. Appl. Genet., 107, 1257, 2003b. Fontana, G.S. et al., Genetic transformation in the grain legume C. arietinum L. (chickpea), Plant

Cell Rept., 12, 194, 1993. Fratini, R. and Ruiz, M.L., A rooting procedure for lentil (Lens culinaris Medik.) and other hypogeous legumes (pea, chickpea and Lathyrus) based on explant polarity, Plant Cell Rept.,21, 726, 2003. Galasso, I. and Pignone, D., Characterization of chickpea chromosomes by banding techniques, Genetic Resources Crop Evol., 39, 115, 1992. Galasso, I. et al., Chromatin characterization by banding techniques, in situ hybridization, and nuclear DNA content in Cicer L. (Leguminosae), Genome, 39, 258, 1996. Gargav, S. and Gaur, P.M., Phylogenetic relationships of perennial chickpea species Cicer

microphyllum (Benth.) with its annual relatives as revealed by allozyme polymorphism, Indian

J. Genet., 61, 218, 2001. Gaur, P.M. and Gour, V.K., Broad-few-leaflets and outwardly curved wings: two new mutants of chickpea, Plant Breed., 122, 192, 2003. Gaur, P.M. and Slinkard, A.E., Genetic control and linkage relations of additional isozyme markers in chickpea, Theor. Appl. Genet., 80, 648, 1990a. Gaur, P.M. and Slinkard, A.E., Inheritance and linkage of isozyme coding genes in chickpea, J.

Hered., 81, 455, 1990b. Gortner, G. et al., Chromosomal localization and distribution of simple sequence repeats and the Arabidopsistype telomere sequence in the genome of Cicer arietinum L., Chromosome Res., 6, 97, 1998.


Gowda, C.L.L. and Gaur, P.M., Global scenario of chickpea research: Present status and future thrusts. In Proceedings of National Symposium on Pulses for Crop Diversification and Natural

Resource Management, December 20–22, 2003, Ali, M., Ed., The Indian Society of Pulses Research and Development, Kanpur, India, 1, 2004. Gupta, P.K. and Bahl, J.R., Cytogenetics and origin of some pulse crops. In Cytogenetics of Crop

Plants, Swaminathan, M.S., Gupta, P.K., and Sinha, U., Eds., Macmillan India, New Delhi, 405, 1983.Gupta, P.K. and Sharma, P.C., Cytogenetics and related aspects in some pulse crops. In Chromosome Engineering in Plants: Genetics, Breeding, Evolution, Part B, Tsuchiya, T. and Gupta, P.K., Eds., Elsevier Science B.V., Amsterdam, The Netherlands, 1, 1991. Gupta, P.K. et al., Molecular markers and their applications in wheat breeding, Plant Breed., 118, 369, 1999. Hajj-Moussa, E. et al., Genetic map of chickpea (Cicer arietinum L.) using morphological and molecular markers. In 4th European Conference on Grain Legumes: Towards the Sustainable

Production of Healthy Foods, Feed and Novel Products, European Association for Grain Legume Research, Paris, 244, 2001. Hamblin, J. et al., The development and status of transgenic pulses in Australia. In Opportunities

for High Quality, Healthy and Added Value Crops to Meet European Demands: Proceedings of

the 3rd European Conference on Grain Legumes, AEP, Paris, 70, 1998. Haq, M.A., Sadiq, M., and Hassan, M., CM 98 (CM 31–1/85): a very high yielding, disease-resistant mutant variety of chickpea, International Chickpea and Pigeonpea Newslt., 6, 9, 1999. Harlan, J.R., Evaluation of wild relatives of crop plants. In Crop Genetic Resources: Conservation

and Evaluation, Holden, J.H.W. and Williams, J.T., Eds., George Allen & Unwin Publishers Ltd., London, 212, 1984. Harlan, J.R. and de Wet, J.M.J., Toward a rational classification of cultivated plants, Taxonomy, 20, 509, 1971. Hassan, A.E., Some seed characters and identification of some Cicer species, Bulletin of Faculty of

Agriculture-University of Cairo, 51, 331, 2000. Haware, M.P., Diseases of chickpea. In The Plant Pathology of Food and Pasture Legumes, Allen, D.J. and Lenne, J.M., Eds., CAB International, Wallingford, U.K., 473, 1998. Haware, M.P. and McDonald, D., Integrated management of botrytis gray mold of chickpea. In Botrytis Gray Mold of Chickpea: Summary Proceedings of the BARI/ICRISAT Working Group

Meeting to Discuss Collaborative Research on Botrytis Gray Mold of Chickpea, Haware, M.P., Faris, D.G. and Gowda, C.L.L., Eds., ICRISAT, Patancheru, India, 1, 1992. Haware, M.P. and McDonald, D., Botrytis gray mold of chickpea. In Recent Advances in Research

on Botrytis Gray Mold of Chickpea: Summary Proceedings of the Second Working Group

Meeting, Haware, M.P., Gowda, C.L.L., and McDonald, D., Eds., ICRISAT, Patancheru, India, 3, 1993. Haware, M.P. and Nene, Y.L., Screening of chickpea for resistance to Botrytis gray mold, International Chickpea Newslt., 6, 17, 1982. Haware, M.P., Narayana Rao, J., and Pundir, R.P.S., Evaluation of wild Cicer species for resistance to four chickpea diseases, International Chickpea Newslt., 27, 16, 1992. Haware, M.P. et al., Integrated management of wilt and root rots of chickpea. In Chickpea in the

Nineties: Proceedings of the Second Int. Workshop on Chickpea Improvement, ICRISAT, Patancheru, India, 129, 1990. Haware, M.P. et al., Multilocation testing of chickpea for field resistance to ascochyta blight, Indian Phytopathol., 48, 289, 1995. Helbaek, H., The plant husbandry at Hacilar. In Excavation at Hacilar, Mellaart, J., Ed., Edinburgh University Press, 189, 1970. Hussain, T. et al., Studies on the expression of marker genes in chickpea, Plant Cell Tissue Organ

Cult., 49, 7, 1997.

Chickpea 259

Idnani, N. and Gaur, P.M., Linkage mapping of two mutant loci controlling leaf necrosis and glabrous shoot in chickpea (Cicer arietinum L.), Indian J. Genet., 63, 45, 2003. Infantino, A., Porta-Puglia, A., and Singh, K.B., Screening wild Cicer species for resistance to fusarium wilt, Plant Disease, 80, 42, 1996. Iruela, M. et al., Phylogenetic analysis in the genus Cicer and cultivated chickpea using RAPD and ISSR markers, Theor. Appl. Genet, 104, 643, 2002. Jaiswal, H.K., Singh, B.D., and Singh, R.M., Improvement of chickpea (Cicer arietinum) through introgression of genes from Cicer reticulatum, Indian J. Agr. Sci, 57, 880, 1987. Jaiwal, P.K., Upadhyaya, S., and Upadhyaya, K.C., Chickpea regeneration and transformation, Current Sci., 80, 1368, 2001. Javadi, F. and Yamaguchi, H., Interspecific relationships of the genus Cicer L. (Fabaceae) based on trnT-F sequences, Theor. Appl. Genet., 109, 317, 2004a. Javadi, F. and Yamaguchi, H., RAPD and seed coat morphology variation in annual and perennial species of the genus Cicer L., Genetic Res. Crop Evol., 51, 783, 2004b. Jayanand, B., Sudarsanam, G., and Sharma, K.K., An efficient protocol for the regeneration of whole plants of chickpea (Cicer arietinum L.) by using axillary meristem explants derived from in vitro-germinated seedlings. In Vitro Cell Dev. Biol. Plant, 39, 171, 2003. Jimenéz-Díaz, R.M., Trapero-Casas, A., and Trapero-Casas, J.L., Races of Fusarium oxysporum f. sp. ciceri infecting chickpea in southern Spain. In Vascular Wilt of Plants, Tjamos, E.C. and Beckman, C, Eds., NATO ASI Series, Vol H28, Springer-Verlag, Berlin, Germany, 515, 1989. Johnston, D.J., Ramanathan, V., and Williamson, B., A protein from immature raspberry fruits which inhibits endopolygalacturonases from Botrytis cinerea and other microorganisms, J. Expt.

Bot., 44, 971, 1993. Kaiser, W.J. et al., Chickpea wilt incited by pea streak carlavirus, Plant Disease, 77, 922, 1993. Kaiser, W.J. et al., Screening of wild Cicer species for resistance to races 0 and 5 of Fusarium

oxysporum f. sp. Ciceris, Plant Disease, 78, 962, 1994. Kaiser, W.J. et al., Growing techniques and conservation of wild perennial Cicer species in the US Pacific northwest, International Chickpea and Pigeonpea Newslt., 4, 7, 1997. Kar, S. et al., Efficient transgenic plant regeneration through Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.), Plant Cell Rept., 16, 32, 1996. Kar, S. et al., Expression of cryIA(C) gene of Bacillus thuringiensis in transgenic chickpea plants inhibits development of pod borer (Heliothis armigera) larvae, Transgenic Res., 6, 177, 1997. Kathiria, K.B. et al., Screening of chickpea genotypes for salinity tolerance during germination and early seedling growth, Gujarat Agriltural Univ. Res. J., 22, 28, 1997. Kaur, S., Chhabra, K.S., and Arora, B.S., Incidence of Helicoverpa armigera (Hubner) on wild and cultivated species of chickpea, Int. Chickpea and Pigeonpea Newsl., 6, 18, 1999. Kazan, K. and Muehlbauer, F.J., Allozyme variation and phylogeny in annual species of Cicer

(Leguminosae), Plant Syst. Evol, 175, 11, 1991. Kazan, K. et al., Inheritance and linkage relationships of morphological and isozyme loci in chickpea (Cicer arietinum L.), Theor. Appl. Genet., 86, 417, 1993. Khanna, V.K. and Maherchandani, N., Comparison of radiosensitivity of different genotypes of chickpea (Cicer arietinum L.), Int. Chickpea Newsl., 4, 4, 1981. Kharbanda, P.D. and Bernier, C.C., Botrytis grey mold of chickpea in Manitoba, Plant Disease

Reporter, 63, 662, 1979. Kharkwal, M.C., Induced mutations in chickpea (Cicer arietinum L.) I. Comparative mutagenic effectiveness and efficiency of physical and chemical mutagens, Indian J. Genet., 58, 159, 1998. Kharkwal, M.C., Jain, H.K., and Sharma, B., Induced mutations for improvement of chickpea, lentil, pea and cowpea. In Improvement of Grain Legume Production Using Induced Mutations,Pullman, WA, 89,1988. Knights, E.J., Brinsmead, R.B., Fordyce, M., Wood, J.A., Kelly, A., and Harden, S., Use of the wild relative Cicer echinospermum in chickpea improvement. In Plant Breeding for the 11th


Millennium: Proc. 12th Australasian Plant Breed. Conf., McComb, J.A., Ed., Australian Plant Breeding Assoc. Inc., Perth, Australia, 145, 2002. Knights, E.J., Southwell, R.J., and Schwinghamer, M.W., Evaluation of wild Cicer species for resistance to phytophthora root rot. In Chickpea Research for the Millennium: Proceedings of

the International Chickpea Conference, Sharma, R.N., Shrivastava, G.K., Rathore, A.L., Sharma, M.L., and Khan, M.A., Eds., Indira Gandhi Agricultural University, Raipur, India, 54, 2003.Krishnamurthy, L. et al., Genetic diversity of drought-avoidance root traits in the mini-core germplasm collection of chickpea, Int. Chickpea and Pigeonpea Newsl., 10, 21, 2003. Krishnamurthy, K.V. et al., Agrobacterium mediated transformation of chickpea (Cicer arietinum)embryo axes, Plant Cell Rept., 19, 235, 2000. Kupicha, F.K., Cicereae Alefeld. In Advances in Legume Systematics Part 1, Polhill, R.M. and Raven, P.H., Eds., Royal Botanic Gardens, Kew, U.K., 382, 1981. Labdi, M. et al., Genetic diversity and phylogenetic relationships among the annual Cicer species as revealed by isozyme polymorphism, Euphytica, 88, 181, 1996. Ladizinsky, G., A new Cicer from Turkey, Notes from the Royal Botanical Garden Edinburgh, 34, 201, 1975. Ladizinsky, G. and Adler, A., The origin of chickpea as indicated by seed protein electrophoresis, Israel J. Bot., 24, 183, 1975. Ladizinsky, G. and Adler, A., Genetic relationships among the annual species of Cicer L., Theor.

Appl. Genet., 48, 197, 1976a. Ladizinsky, G. and Adler, A., The origin of chickpea Cicer arietinum L., Euphytica, 25, 211, 1976b.Lateef, S.S. and Pimbert, M.P., The search for host resistance to Helicoverpa armigera. In

Chickpea and Pigeonpea at ICRISAT, Proceedings of the First Consultative Group Meeting on

Host Selection Behaviour of Helicoverpa armigera, ICRISAT, Patancheru, India, 1990. Lather, V.S., Promising chickpea ideotype for higher plant density, Intl. Chickpea Newsl., 7, 26, 2000.Lavanya, M. et al., Genetic transformation of pigeonpea to evaluate insecticidal genes for resistance to the legume pod borer. In 7th International Congress of Plant Molecular Biology,www.ispmb2003.com/abst, Barcelona, 2003. Leport, L. et al., Physiological responses of chickpea genotypes to terminal drought in a Mediterranean type environment, Europeon J. Agron., 11, 279, 1999. Lülsdorf, M. et al., Development of microspore culture technology for the production of doubled-haploid lines of chickpea (Cicer arietinum L.). In Proceedings of the North American Pulse

Improvement Association Meeting, Fargo, ND, 17, 2001. Malhotra, R.S. et al., Genetic improvement and agronomic management of chickpea with emphasis on the Mediterranean region. In Adaptation of Chickpea in the West Asia and North Africa

Region, Saxena, N.P., Saxena, M.C., Johansen, C., Virmani, S.M., and Harris, H., Eds., ICRISAT, Patancheru, India and ICARDA, Aleppo, Syria, 217, 1996. Malhotra, R.S. et al., Cicer species—conserved resources, priorities for collection and future prospects. In Linking Research and Marketing Opportunities for Pulses in the 21st Century,Knight, R., Ed., Kluwer Academic Publishers, Dordrecht, The Netherlands, 603, 2000. Malhotra, R.S. et al., Registration of ILC 10765 and ILC 10766 chickpea germplasm lines resistant to cyst nematode, Crop Sci., 42, 1756, 2002. Mallikarjuna, N., Ovule and embryo culture to obtain hybrids from interspecific incompatible pollinations in chickpea, Euphytica, 110, 1, 1999. Mallikarjuna, N., Prospects of using Cicer canariense for chickpea improvement, Int. Chickpea

and Pigeonpea Newsl., 8, 23, 2001. Mathur, D.S. and Lal, S.K., Chemical induction of male sterility in chickpea, Indian J. Genet., 59, 3, 379, 1999.

Chickpea 261

Mayer, M.S. et al., Development of a DNA marker for fusarium wilt resistance in chickpea, Crop

Sci., 37, 1625, 1997. Micke, A., Genetic improvement of food legumes in developing countries by mutation induction. In World Crops: Cool Season Food Legumes, Summerfield, R.J., Ed., Kluwer Academic Publisher, Dordrecht, The Netherlands, 1031, 1988. Millan, T. et al., Markers associated with ascochyta blight resistance in chickpea and their potential in markerassisted selection, Field Crops Res., 84, 373, 2003. Morgan, J.M., Rodriguez, M.B., and Knights, E.J., Adaptation to water deficit in chickpea breeding lines by osmoregulation: Relationship to grain yields in the field, Field Crop Res., 27, 61, 1991. Muehlbauer, F.J., personal communication, 2004. Muehlbauer, F.J., Kaiser, W.J., and Simon, C.J., Potential for wild species in cool season food legume breeding, Euphytica, 73, 109, 1994. Muehlbauer, F.J. et al., Population improvement in pulse crops: An assessment of methods and techniques. In World Crops: Cool Season Food Legumes, Summerfield, R.J., Ed., Kluwer Academic Publisher, Dordrecht, The Netherlands, 943, 1988. Narvel, J.M. et al., Development of multiple sets of simple sequence repeat DNA markers covering the soybean genome, Mol. Breed., 6, 175, 2000. Nene, Y.L. and Haware, M.P., Screening chickpea for resistance to wilt, Plant Disease, 64, 379, 1980.Nene, Y.L., Haware, M.P., and Reddy, M.V., Chickpea diseases: Resistance-screening techniques, Information Bulletin No. 10, International Crops Research for the Semi-Arid Tropics, Patancheru, India, 1981. Nene, Y.L., Sheila, V.K., and Sharma, S.B., A World List of Chickpea and Pigeonpea Pathogens,5th edition, ICRISAT, Patancheru, India, 1996. Nguyen, T.T. et al., Genetic diversity estimates in Cicer using AFLP analysis, Plant Breed., 123, 173, 2004. Nozzolillo, C, Seedling morphology and anatomy of eight Cicer species and their taxonomic value, Can. J. Bot., 63, 1, 1985. Ocampo, B., Robertson, L.D., and Singh, K.B., Variation in seed protein content in the annual wild Cicer species, J. Sci. Food Agril., 78, 220, 1998. Ocampo, B. et al., Karyotype analysis in the genus Cicer, J. Genet. Breed., 46, 229, 1992. Ohri, D., Cytology of Cicer songaricum Steph. ex DC, a wild relative of chickpea, Genetic

Resources and Crop Evolution, 46, 111, 1999. Ohri, D. and Pal, M., The origin of chickpea (Cicer arietinum L.): karyotype and nuclear DNA amount, Heredity, 66, 367, 1991. Oram, R.N. et al., Isozyme similarity and genetic differences in morphology between Hyprosola, a high yielding, high protein mutant of chickpea (Cicer arietinum L.), and its parental cultivar, Environmental and Expt. Bot., 27, 455, 1987. Patil, P.B. et al., Variation in the ribosomal RNA units of the genera Lens and Cicer, Euphytica, 83, 33, 1995. Pfaff, T. and Kahl, G., Mapping of gene-specific markers on the genetic map of chickpea (Cicer

arietinum L.), Mol. Genet. Genomics, 269, 243, 2003. Phillips, J.C., A distinct race of chickpea wilt in California, Int. Chickpea Newsl., 18, 19, 1988. Porta-Puglia, A. and Infantino, A., The search for sources of resistance to Fusarium wilt in wild Cicer species: a synthesis of the Italian experience, Pakistan J. Phytopathol., 9, 26, 1997. Pundir, R.P.S., personal communication, 1994. Pundir, R.P.S. and Mengesha, M.H., Cross compatibility between chickpea and its wild relative, Cicer echinospermum Davis., Euphytica, 83, 241, 1995. Pundir, R.P.S. and Reddy, G.V., Two new traits—open flower and small leaf in chickpea Cicer

arietinum L.), Euphytica, 102, 357, 1988. Pundir, R.P.S. and van der Maesen, L.J.G., Interspecific hybridization in Cicer., Int. Chickpea

Newsl., 8, 4, 1983.


Pundir, R.P.S., Mengesha, M.H., and Reddy, G.V., Interspecific hybridization in Cicer., Int.

Chickpea Newsl., 26, 6, 1992. Pundir, R.P.S., Mengesha, M.H., and Reddy, G.V., Morphology and cytology of Cicer canariense,a wild relative of chickpea, Euphytica, 69, 73, 1993. Pundir, R.P.S., Rao, N.K., and van der Maesen, L.J.G., Distribution of qualitative traits in the world germplasm of chickpea (Cicer arietinum L.), Euphytica, 34, 697, 1985. Pundir, R.P.S., Reddy, K.N., and Mengesha, M.H., ICRISAT Chickpea Germplasm Catalog: Evaluation and Analysis, ICRISAT, Patancheru, India, 1988. Rajesh, P.N. et al., Identification of an STMS marker for the double-podding gene in chickpea, Theor. Appl Genet., 105, 604, 2002a. Rajesh, P.N. et al., Molecular mapping and characterization of an RGA locus RGAPotkinl-2171 in chickpea, Euphytica, 128, 427, 2002b. Rajesh, P.N. et al., Genetic relationships among annual and perennial species of Cicer using inter simple sequence repeat (ISSR) polymorphism, Euphytica, 129, 15, 2003. Rajesh, P.N. et al., Construction of a Hind III bacterial artificial chromosome library and its use in identification of clones associated with disease resistance in chickpea, Theor. Appl. Genet., 108, 663, 2004. Rakshit, S. et al., DAF marker tightly linked to a major locus for ascochyta blight resistance in chickpea (Cicer arietinum L.), Euphytica, 132, 23, 2003. Ratnaparkhe M.B., Tekeoglu, M., and Muehlbauer, F.J., Inter simple sequence repeat (ISSR) polymorphisms are useful for finding markers associated with disease resistance gene clusters, Theor. Appl. Genet., 97, 515, 1998a. Ratnaparkhe, M.B. et al., Inheritance of inter-simple-sequence-repeat polymorphisms and linkage with a fusarium wilt resistance gene in chickpea, Theor. Appl. Genet., 96, 348, 1998b. Reddy, V.D. and Reddy, G.M.. In vivo production of haploids in chickpea (Cicer arietinum L.), J.

Genet. Breed., 51, 1, 29, 1997. Reddy, M.V. and Singh, K.B., Evaluation of a world collection of chickpea germplasm accessions for resistance to Ascochyta blight, Plant Disease, 68, 900, 1984. Reddy, M.V., Singh, K.B., and Malhotra, R.S., Multilocation evaluation of chickpea germplasm and breeding lines for resistance to Ascochyta blight, Phytopathology Mediterranean, 31, 59, 1992.Robertson, L.D., Ocampo, B., and Singh, K.B., Morphological variation in wild annual Cicerspecies in comparison with the cultigen, Euphytica, 95, 309, 1997. Robertson, L.D., Singh, K.B., and Ocampo, B., A catalog of annual wild Cicer species, ICARDA, Aleppo, Syria, 1995. Robertson, L.D. et al., Useful genetic diversity in germplasm collections of food and forage legumes from West Asia and North Africa, Genetic Resources Crop Evolution, 43, 447, 1996. Rubiales, D., Alcantara, C., and Sillero, J.C., Variation in resistance to Orobanche crenata in species of Cicer. Weed Res., 44, 27, 2004.Rubio, J. et al., Two genes and linked RAPD markers involved in resistance to Fusarium

oxysporum f. sp ciceris race 0 in chickpea, Plant Breed., 122, 188, 2003. Sagare, A.P. and Krishnamurthy, K.V., Protoplast regeneration in chickpea, Cicer arietinum L., Indian J. Expl. Biol., 29, 930, 1991. Saini, H.S., Weder, J.K.R, and Knights, E.J., Inhibitor activities of chickpeas (Cicer arietinum L.) against bovine, porcine and human trypsin and chymotrypsin, J. Sci. Food and Agril., 60, 287, 1992.Santra, D.K. et al., Identification and mapping of QTLs conferring resistance to Ascochyta blight in chickpea, Crop Sci., 40, 1606, 2000. Saraf, C.S. et al., Biological nitrogen fixation and residual effects of winter grain legumes in rice and wheat cropping systems of the Indo-Gangetic plain. In Residual Effects of Legumes in Rice

and Wheat Cropping Systems of the Indo-Gangetic Plain, Kumar Rao, J.V.D.K., Johansen, C., and Rego, T.J., Eds., Oxford and IBH Publishing Co. Pvt. Ltd., New Delhi, 14, 1998.

Chickpea 263

Saxena, N.P., Management of drought in chickpea—a holistic approach. In Management of

Agricultural Drought—Agronomic and Genetic Options, Saxena, N.P., Ed., Oxford & IBH Publishing Co. Pvt. Ltd., New Delhi, 103, 2003. Saxena, N.P. and Johansen, C., Chickpea ideotypes for genetic enhancement of yield and yield stability in south Asia. In Chickpea in the Nineties: Proceedings of the Second International

Workshop on Chickpea Improvement, ICRISAT, Patancheru, India, 81, 1990. Sedgley, R.H., Siddique, K.H.M., and Walton, G.H., Chickpea ideotypes for Mediterranean environments. In Chickpea in the Nineties: Proceedings of the Second International Workshop

on Chickpea Improvement, ICRISAT, Patancheru, India, 87, 1990. Senthil, G., Williamson, B., Dinkins, R.D., and Ramsay, G., An efficient transformation system for chickpea (Cicer arietinum L.), Plant Cell Rep., 23, 297, 2004. Sethi, S.C., Male sterility, Int. Chickpea Newsl., 1, 4, 1979. Sharma, H.C. et al., Identification of resistance to Helicoverpa in wild species of chickpea. In Plant

Breeding for the 11th Millennium: Proceedings of the 12th Australian Plant Breeding

Conference, McComb, J.A., Ed., Australian Plant Breeding Assoc., Perth, Australia, 294, 2002. Sharma, H.C. et al., Host plant resistance to pod borer, Helicoverpa armigera in chickpea. In Chickpea Research for the Millennium: Proceedings of the International Chickpea Conference,Sharma, R.N., Shrivastava, G.K., Rathore, A.L., Sharma, M.L., and Khan, M.A., Eds., Indira Gandhi Agricultural University, Raipur, India, 118, 2003. Sharma, H.C., Bhagwat, M.P., Pampapathy, G., Sharma, J.P., and Ridshill-Smith, T.J., Perennial wild relatives of chickpea as potential sources of resistance to Helicoverpa armigera. Genetic

Res. Crop Evol., www.kluweronline.com/issn/0925–9864, (In Press, Pre-publication date: Sep 3, 2004), 2004. Sharma, K.D. et al., Molecular mapping of Fusarium oxysporum f. sp. ciceris race 3 resistance gene in chickpea, Theor. Appl. Genet., 108, 1243, 2004. Sharma, K.K., personal communication, 2003. Sharma, K.K. and Ortiz, R., Program for the application of genetic transformation for crop improvement in the semi-arid tropics, in vitro Cellular Developmental Biol.-Plant, 36, 83, 2000. Sharma, P.C. et al., The potential of microsatellites for hybridisation and polymerase chain reaction-based DNA fingerprinting of chickpea (Cicer arietinum L.) and related species, Electrophoresis, 16, 1755, 1995. Sheila, V.K. et al., Interspecific hybridisation between Cicer arietinum and wild Cicer species, Int.

Chickpea Newsl., 27, 11, 1992. Siddique, K.H.M. and Sykes, J., Pulse production in Australia past, present and future, Australian

J. Exptl Agricult., 37, 103, 1997. Siddique, K.H.M. et al., Adaptation of chickpea (Cicer arietinum L.) and faba bean (Vicia faba L.) to Australia. In Linking Research and Marketing Opportunities for Pulses in the 21st Century,Knight, R., Ed., Kluwer Academic Publishers, Dordrecht, The Netherlands, 289, 2000. Simon, C.J. and Muehlbauer, F.J., Construction of a chickpea linkage map and its comparison with maps of pea and lentil, J. Hered., 88, 115, 1997. Singh, A., Singh, N.P., and Asthana, A.N., Genetic potential of wide crosses in chickpea, Legume

Res., 22, 19, 1999a. Singh, B.D. et al., Isolation of early-flowering recombinants from the interspecific cross between Cicer arietinum and C. reticulatum, Int. Chickpea Newsl., 11, 14, 1984a. Singh, K.B., Winter chickpea: Problems and potential in the Mediterranean region. In Present

Status and Future Prospect of Chickpea Crop Production and Improvement in the

Mediterranean Countries, Saxena, M.C., Cubero, J.I., and Wery, J., Eds., Options Mediterranéennes, Serie A: Seminaires Mediterranéenes: No. 9, CIHEAM, Zaragoza, Spain, 1990.Singh, K.B. and Ocampo, B., Interspecific hybridization in annual Cicer species, J. Genet. Breed.,47, 199, 1993.


Singh, K.B. and Ocampo, B., Exploitation of wild Cicer species for yield improvement in chickpea, Theor. Appl. Genet., 95, 418, 1997. Singh, K.B. and Reddy, M.V., Advances in disease resistance breeding in chickpea, Adv. Agron.,45, 191, 1991. Singh, K.B. and Reddy, M.V., Sources of resistance to ascochyta blight in wild Cicer species, Netherlands J. Plant Pathol., 99, 163, 1993. Singh, K.B. and Weigand, S., Identification of resistant sources in Cicer species to Liriomyza

cicerina, Genetic Resources and Crop Evolution, 41, 75, 1994. Singh, K.B., Malhotra, R.S., and Saxena, M.C., Sources for tolerance to cold in Cicer species, Crop

Sci., 30, 1136, 1990. Singh, K.B., Malhotra, R.S., and Saxena, M.C., Additional sources of tolerance to cold in cultivated and wild Cicer species, Crop Sci., 35, 1491, 1995. Singh, K.B., Ocampo, B., and Robertson, L.D., Diversity for abiotic and biotic stress resistance in the wild annual Cicer species, Genetic Resources Crop Evolution, 45, 9, 1998. Singh, K.B., Reddy, M.V., and Nene, Y.L., International testing of chickpeas for resistance to ascochyta blight, Plant Disease, 68, 782, 1984b. Singh, K.B. et al., Reaction of wild Cicer spp. lines to Heterodera ciceri, Nematology

Mediterranean, 17, 113, 1989. Singh, K.B. et al., Current status and future strategy in breeding chickpea for resistance to biotic and abiotic stresses, Euphytica, 73, 137, 1994. Singh, K.B. et al., Registration of ILWC 292, a chickpea cyst nematode-resistant germplasm of Cicer reticulatum Ladiz., Crop Sci., 36, 1421, 1996. Singh, N.B. and Singh, R.A., Screening for alkalinity and salinity tolerance in chickpea, Int.

Chickpea Newsl., 11, 25, 1984. Singh, N.P., Singh, A., and Asthana, A.N., Studies on inter-specific crossability barriers in chickpea, Indian J. Pulses Res., 12, 13, 1999b. Singh, R.P. and Singh, B.D., Recovery of rare interspecific hybrids of gram Cicer arietinum×C.

cuneatum L. through tissue culture, Current Sci., 58, 874, 1989. Singh, S.D., personal communication, 2003. Singh, U. and Pundir, R.P.S., Amino acid composition and protein content of chickpea and its wild relatives, Int. Chickpea Newsl., 25, 19, 1991. Sommers, D.A., Samac, D.A., and Olhoft, P.M., Recent advances in legume transformation, Plant

Physiol., 131, 892, 2003. Staginnus, C. et al., Molecular structure and chromosomal localization of major repetitive DNA families in the chickpea (Cicer arietinum L.) genome, Plant Mol. Biol., 39, 1037, 1999. Staginnus, C. et al., A PCR-based assay to detect En/Spm-like transposon sequences in plants, Chromosome Res., 9, 591, 2001. Stamigna, C., Crinò, P., and Saccardo, F., Wild relatives of chickpea: multiple disease resistance and problems to introgression in the cultigen, J. Genet. Breed., 54, 213, 2000. Stamigna, C. et al., Multiple resistance to diseases in wild relatives of chickpea (Cicer arietinumL.). In Opportunities for High Quality, Healthy and Added Value Crops to Meet European

Demands: Proceedings of the 3rd European Conference on Grain Legumes, Valladolid, Spain, 221, 1998. Sudupak, M.A., Inter and intra-species Inter Simple Sequence Repeat (ISSR) variations in the genus Cicer., Euphytica, 135, 229, 2004. Sudupak, M.A. and Kence, A., Genetic relationships among perennial and annual Cicer species growing in Turkey as revealed by allozymes, Genetic Res. Crop Evol., 51, 241, 2004. Sudupak, M.A., Akkaya, M.S., and Kence, A., Analysis of genetic relationships among perennial and annual Cicer species growing in Turkey using RAPD markers, Theor. Appl. Genet., 105, 1220, 2002. Sudupak, M.A., Akkaya, M.S., and Kence, A., Genetic relationships among perennial Cicer species growing in Turkey assessed by AFLP fingerprinting, Theor. Appl. Genet, 108, 937, 2004.

Chickpea 265

Swamy, A.V.S.R. and Khanna, V.K., Pollen grain germination and pollen tube growth following interspecific pollination in Cicer, SABRAO J., 23, 137, 1991. Tayyar, R.I. and Waines, J.G., Genetic relationships among annual species of Cicer (Fabaceae) using isozyme variation, Theor. Appl. Genet, 92, 245, 1996. Tayyar, R.I., Federici, C.V., and Waines, G.J., Natural outcrossing in chickpea (Cicer arietinumL.), Crop Sci., 36, 203, 1995. Tayyar, R.I., Lukaszewski, A.J., and Waines, J.G., Chromosome banding patterns in the annual species of Cicer, Genome, 37, 656, 1994. Tekeoglu, M., Rajesh, P.N., and Muehlbauer, F.J., Integration of sequence tagged microsatellite sites to the chickpea genetic map, Theor. Appl. Genet., 105, 847, 2002. Tewari-Singh, N. et al., Use of a herbicide or lysine plus threonine for non-antibiotic selection of transgenic chickpea, Plant Cell Rept., 22, 576, 2004. Than, H., Thein, M.M., and Myint, S.S., Relationship among Rhizoctonia bataticola isolates in rice-based cropping systems based on colony fusion types, Int. Chickpea Newsl., 25, 29, 1991. Toker, C., Preliminary screening and selection for cold tolerance in annual wild Cicer species, Genetic Resources Crop Evol, www.kluweronline.com/issn/0925–9864, (In Press), 2004. Tullu, A. et al., Inheritance and linkage of a gene for resistance to race 4 of fusarium wilt and RAPD markers in chickpea, Euphytica, 102:227, 1998. Tullu, A. et al., A second gene for resistance to race 4 of Fusarium wilt in chickpea and linkage with a RAPD marker, Euphytica, 109, 43, 1999. Tuwafe, S. et al., Inheritance and geographical distribution of allozyme polymorphisms in chickpea (Cicer arietinum L.), J. Hered., 79, 170, 1988. Udupa, S.M. et al., Narrow genetic variability in Cicer arietinum L. as revealed by RFLP analysis, J. Plant Biochem. Biotechnol., 2, 83, 1993. Upadhyaya, H.D. and Ortiz, R., A mini core subset for capturing diversity and promoting utilization of chickpea genetic resources in crop improvement, Theor. Appl. Genet., 102, 1292, 2001.Upadhyaya, H.D., Bramel, P.J., and Singh, S., Development of a chickpea core subset using geographic distribution and quantitative traits, Crop Sci., 41, 206, 2001. Vairinhos, F. and Murray, D.R., The seed proteins of chickpea; comparative studies of Cicer

arietinum, C. reticulatum and C. echinospermum (Leguminosae), Plant Syst. Evol, 142, 11, 1983.Valarik, M. et al., High-resolution FISH on super-stretched flow-sorted plant chromosomes, The

Plant J., 37, 940, 2004. van der Maesen, L.J.G., Origin, history and taxonomy of chickpea. In The Chickpea, Saxena, M.C. and Singh, K.B., Eds., CAB International, Cambridge, U.K., 11, 1987. van der Maesen, L.J.G. and Pundir, R.P.S., Availability and use of wild Cicer germplasm, FAO/IBPGR Plant Genetic Resources Newslt., 57, 19, 1984. van Dorrestein, B., Baum, M., and Malhotra, R.S., Interspecific hybridisation between cultivated chickpea (Cicer arietinum L.) and the wild annual species C. judaicum Boiss, C. pinnatifidumJaub. & Sp. and C. bijugum K.H. Rech. In Opportunities for High Quality, Healthy and Added

Value Crops to Meet European Demands: Proceedings of the 3rd European Conference on

Grain Legumes, Valladolid, Spain, 362, 1998. van Rheenen, H.A., Chickpea breeding—progress and prospects, Plant Breed. Abstr., 61, 997, 1991.van Rheenen, H.A., Pundir, R.P.S., and Miranda, J.H., How to accelerate the genetic improvement of a recalcitrant crop species such as chickpea, Current Sci., 65, 10, 1993. van Rheenen, H.A. et al., Polygon breeding: old hat or new trick?, Int. Chickpea Newsl., 25, 1991. van Rheenen, H.A. et al., Breeding for resistance to soil-borne diseases in chickpea. In Disease

Resistance Breeding in Chickpea, Singh, K.B. and Saxena, M.C., Eds., ICARDA, Aleppo, Syria, 1992.


Verma, M.M., Ravim M., and Sandhu, J.S., Characterization of the interspecific cross Cicer

arietinum L.× C. judaicum (Boiss), Plant Breed., 114, 549, 1995. Verma, M.M. et al., Crossability studies in different species of Cicer (L.), Crop Improvement, 17, 179, 1990. Vlacilova, K. et al., Development of flow cytogenetics and physical genome mapping in chickpea (Cicer arietinum L.), Chromosome Res., 10, 695, 2002. Williams, P.C. and Singh, U., The Chickpea—Nutritional quality and the evaluation of quality in breeding programmes. In The Chickpea, Saxena, M.C. and Singh, K.B., Eds., ICARDA, Aleppo, Syria, 329, 1987. Winter, P. et al., Characterization and mapping of sequence tagged microsatellite sites in the chickpea (Cicer arietinum L.) genome, Mol. General Genet., 262, 90, 1999. Winter, P. et al., A linkage map of the chickpea (Cicer arietinum L.) genome based on recombinant inbred lines from a C. arietinum×C. reticulatum cross: Localization of resistance genes for fusarium wilt races 4 and 5, Theor. Appl. Genet., 101, 1155, 2000. Zohary, D. and Hopf, M., Pulses. In Domestication of Plants in the Old World: The Origin and

Spread of Cultivated Plants in West Asia, Europe, and the Nile Valley, 3rd Edition, Oxford University Press, New York, 108, 2000.

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